JP6753862B2 - Improved ionization of gas samples - Google Patents

Description

Mutual reference of related applications This application is written in UK Patent Application No. 1503876.3 filed on March 6, 2015, UK Patent Application No. 1503864.9 filed on March 6, 2015, October 2015. UK Patent Application No. 15183699.2 filed on 16th March, UK Patent Application 1503877.1 filed on 6th March 2015, UK Patent Application 1503876 filed on 6th March 2015 .2, UK Patent Application No. 1503863.1 filed on March 6, 2015, UK Patent Application No. 1503878.9 filed on March 6, 2015, filed on March 6, 2015 Priority and interests are claimed from the UK Patent Application No. 1503879.7 filed and the UK Patent Application No. 1516039.9 filed on September 9, 2015. The entire contents of these applications are incorporated herein by reference.

The present invention relates generally to mass spectrometry, and more specifically to devices for improving sample ionization. Embodiments include rapid evaporative ionization mass spectrometry, mass spectrometry, ion mobility analysis, hybrid ion mobility mass spectrometry, rapid evaporation ionization mass spectrometry (“REIMS”) method, mass spectrometry method, ion mobility method, hybrid. Ion Mobility Mass Spectrometry Methods, Electrosurgical Methods, and Electrosurgical Devices.

Rapid Evaporation Ionized Mass Spectrometry (“REIMS”) is a technique recently developed for real-time identification of substances, eg, for identification of biological tissue during surgical intervention. REIMS analysis of biological tissue is similar to matrix-assisted laser desorption ionization (“MALDI”), secondary ion mass spectrometry (“SIMS”), and desorption electrospray ionization (“DESI”) imaging, as well as high histology and It has been shown to obtain a profile of phospholipids with histopathological specificity.

Combining REIMS technology with a portable sampling device results in iKnife sampling technology, which can provide intraoperative tissue identification. This technique can help surgeons minimize the amount of healthy tissue that the surgeon removes during surgery, such as tumors, but it makes the target tissue more effective by providing useful information to remove the target tissue. Can be excised. The iKnife sampling technique can also be used by non-surgical workers in a non-surgical manner to separate the target object from the substrate in vitro.

In a known iKnife sampling system, the mass spectrometry signal is acquired by the substrate receiving an alternating current at a radio frequency, thereby localizing Joule heat and crushing the cells with the desorption of charged and neutral particles. The resulting aerosol (eg, "surgical smoke") is introduced directly into the atmospheric interface of the atmospheric ionization mass spectrometer for online mass spectrometry. Aerosols contain a sufficient number of ionized molecules so that mass spectrometry can directly fingerprint biological tissue.

Neutral molecules in the sample ionized after evaporation may be used to increase ion yield. In this regard, electrospray and corona discharge post-ionization methods were tested. Secondary electrospray ionization, molten droplet electrospray ionization, and extractive electrospray ionization have been used to increase ion yields. These three techniques are similar in that the charged solvent droplets are melted with the aerosol particles in the gas phase and the resulting molten droplets undergo an electrospray-like ionization process. However, these techniques are due to the brittleness of the electrospray installation, the DESI-like phenomenon, the constraints on solvent-based electrospray, and the sample carry-over effect brought about by the flow rate, and the high pressure involved in these techniques. It is bothered by consideration of patient safety in the intervening environment.

It is also possible to enhance ionization by promoting the collision of aerosol particles with the collision surface in the vacuum region of the mass spectrometer. A method for generating collision ions has been developed and disclosed in International Publication No. 2013/098642 (Medimass), in which aerosol particles enter the analyzer at an atmospheric interface and accelerate into the vacuum region of a free jet analyzer. Will be done. The aerosol particles accelerated by the free jet are then directed to the collision surface, increasing the ion yield of the REIMS method.

But despite this promotion, many problems still remain. For example, the ionization yield for this technique remains relatively low. The use of electrosurgical diathermy in a coagulation manner may also result in a lack of ionization or suppression of sample ion formation. Also, when tissues with high triglyceride content such as triglycerides or diglycerides (eg, adipose tissue or breast tissue) are cleaved, ionization may be lacking.

Therefore, it is desired to provide improved methods and devices.

According to the first aspect, it is mass spectrometry and / or ion mobility analysis.
Providing samples and
Feeding the sample with the matrix compound such that the sample is diluted or dissolved in the matrix or forms a first cluster in the matrix.
A method is provided that comprises colliding the first cluster or first droplet of the diluted or dissolved matrix with a collision surface.

WO 2013/098642 describes how to generate and analyze aerosol samples. The aerosol sample produced comprises droplets covered with polar lipids. During the analysis, the droplet is accelerated by the expansion of the free jet within the atmospheric inlet of the mass spectrometer so that the droplet achieves high speed and then hits the collision surface. As a result, gas ions of polar lipid molecules are generated. However, the ionization yield of this technique is relatively low.

The ion yield of this method is relatively low due to the low conversion rate at which the droplets become individual molecular species, mainly due to the strong intermolecular bonds between the sample molecules.

According to embodiments of the invention, the specimen is diluted by or dissolved in the matrix. For example, the specimen may be provided in the form of droplets, aerosols, or liquids, may be melted or combined with the matrix, or may be dissolved in the matrix. The matrix may be in the form of droplets, solids, aerosols, or liquids upon contact with the specimen. Diluting or dissolving the sample in the matrix may substantially remove or reduce the intermolecular bonds between the sample molecules. Therefore, when a diluted or dissolved sample droplet subsequently collides with the collision surface, the sample droplet breaks into smaller droplets, and any given smaller droplet is in the absence of a matrix. It tends to contain fewer sample molecules than it should contain. This makes the generation of sample ions more efficient.

Most of the sample ionization is believed to be due to the ion dissociation of the sample in the solution phase due to the interaction with the counterions present in the sample being analyzed. Diluting or dissolving the sample in the matrix reduces the concentration of the sample in each droplet, promotes ion dissociation in the solution phase, and ultimately increases the proportion of sample ionized. .. Therefore, any matrix that dissolves or dilutes the sample may be used.

According to the various embodiments described herein, the step of causing the first cluster or droplet to collide with the collision surface is to force the first cluster or droplet into a plurality of second smaller clusters or liquids. You may crush it into drops. However, other embodiments are intended to use other forms of dissociation or disintegration of droplets, such as laser irradiation, ultrasonic energy, glow discharge, or photoionization.

The step of providing the specimen may include providing the specimen in the form of a gas phase specimen, aerosol, vapor, smoke, or liquid, and / or the matrix may be fed to the specimen, but the specimen. Is in the form of gas phase specimens, aerosols, vapors, smoke, solids, or liquids.

In this method, a first droplet may be formed by feeding a matrix to a sample, but the sample is in the form of an aerosol, vapor, or smoke, and the matrix is in the form of an aerosol, vapor, or solid. is there.

The step of feeding the matrix to the sample may include feeding the matrix molecules to the sample and mixing the matrix with the sample, the matrix being in the gas phase or in an aerosol, vapor, or the like. Or it is in solid form.

A mixture of the sample and the matrix is either dissolved in the liquid matrix or diluted by the liquid matrix as the gas phase matrix cools and condenses into a liquid and the sample forms the first droplet. As such, it may be transferred from the high pressure region to the low pressure region.

Alternatively, the matrix may be supplied to the sample as a liquid or may be intermixed with the sample.

If the specimen and / or matrix is in the form of a liquid, the specimen and / or matrix may be converted into droplets or vapors, for example by spraying or spraying. For example, if the sample and matrix are mixed as a liquid, the mixture may then be converted into a first sample droplet, eg, by spraying or spraying.

The matrix may first be supplied as a solid (eg, powder), sublimated or melted and evaporated to form a vapor or gas phase that intermixes with the sample. For example, a solid matrix may be mixed with the sample. When the specimens are mixed in liquid form, the mixture may be dried, for example crystals may be formed. The mixture may then be heated to sublimate and / or evaporate the matrix and / or specimen. Examples of suitable matrices include the MALDI matrix and other matrices such as coumarin, 9-aminoacridine, 2,5-dihydroxybenzoic acid, THAP, CHCA, and quercetin.

The matrix comprises (i) a solvent for the sample, (ii) an organic solvent, (iii) a volatile compound, (iv) a polar molecule, (v) water, (vi) one or more alcohols, (vii). Methanol, (viii) ethanol, (ix) isopropanol, (x) acetone, (xi) acetonitrile, (xii) 1-butanol, (xiii) tetrahydrofuran, (xiv) ethyl acetate, (xv) ethylene glycol, (xvi) It may be selected from the group consisting of dimethyl sulfoxide, (xvii) aldehyde, (xvii) ketone, (xiv) non-polar molecule, (xx) hexane, (xxx) chloroform, (xxxi) butanol, and (xxiii) propanol. ..

By way of example, for specimens with polar lipids, the low molecular weight alcohol may be used as a matrix (eg methanol, ethanol, isopropanol) or a ketone (eg acetone). These matrices have been shown to enhance species ionization, otherwise low intensity and no vapor in the matrix has been detected.

Proton matrix solvents may be used, for example, for the analysis of lipids or triglycerides. Alternatively, aprotonic or aproton matrix solvents may be used, for example, for protein analysis.

The mixture of sample and matrix may be a homogeneous mixture or a heterogeneous mixture.

The matrix and / or the sample may be doped with one or more additives to increase the solventization of the sample in the matrix or to increase the ionization of the sample.

The matrix or specimen may be doped with acidic or basic additives. For example, the matrix may be doped with formic acid, diethylamine.

The matrix may result in derivatization of the specimen. For example, the matrix may result in the derivatization of cholesterol or steroids within the specimen. This may indicate an ionized specimen more easily.

The method is to add a calibrator to the matrix and / or sample, or to select a compound in the matrix and / or sample as the calibrator, and to calibrate the method of mass spectrometry and / or ion mobility analysis. It may include using the body. This is particularly beneficial for ambient ionization, such as the REIMS technique, and it can be difficult to introduce a calibrator according to conventional techniques.

The method may include analyzing the rock mass species, for example by adding the rock mass species to the matrix and / or specimen. Rockmass may then be used to correct the mass (s) of the specimen (s). Alternatively or additionally, the method may include analyzing the lock mobility species, eg, by adding the lock mobility species to the matrix and / or specimen. The lock mobility species may then be used to correct the mobility or drift time (s) of the specimen (s).

The step of colliding the first cluster or first droplet with the collision surface may include accelerating the first cluster or first droplet to the collision surface.

The step of accelerating the first cluster or first droplet to the collision surface may include using a pressure differential to accelerate the first cluster or first droplet to the collision surface. Good.

A vacuum pump is used to create a pressure difference between the two regions in order to accelerate the first cluster or first droplet to the collision surface between the first region and the second region. You may. The device may include a mass spectrometer and / or an ion mobility analyzer having an atmospheric interface located between the first region and the second region, the second region being in a vacuum pump. It may be provided with a vacuum chamber that is connected and houses the collision surface.

The combination of gas flow and matrix flow between the first region and the second region (eg, entering the analyzer) may atomize the matrix, producing droplets of appropriate diameter / volume. The spray stream may generate shear forces on the matrix, producing droplets that enter the sample transfer tube.

The method may include evaporating the matrix from a sample in a second cluster or droplet, resulting in free sample ions separated from the matrix.

The step of colliding the first cluster or the first droplet with the collision surface may evaporate the matrix from the sample by converting the kinetic energy of the sample and matrix into heat.

The step of colliding the first cluster or the first droplet may produce a second smaller cluster or droplet, at least a portion of which may have only a single sample molecule therein. This improves the ionization process.

The permittivity of the matrix may be high enough that the solvation of the sample in the matrix involves ion dissolution and results in the solvating ions of the sample present in the condensed phase. In these cases, the collision with the collision surface is likely to produce solvated ions in the gas phase, which are ultimately formed by deprotonation (in negative ion mode, ie [MH] ⁻ ). It may give rise to ionized ions, ions formed by protonation (in positive ion mode, i.e. [M + H] ⁺ ), and / or molecular ions. Alternatively or additionally, the ions may be formed by any one of dehydration, deamidation, or alkali metal adduct.

The method may include ionizing the sample or sample ion downstream of the collision surface. Arbitrarily, the ionization is corona discharge ionization source, reagent ionization source, photoionization source, chemical ionization source, electric shock (“EL”) ionization source, electric field ionization (“FI”) source, electric field desorption (“FD”). ”) Ionization source, inductively coupled plasma (“ICP”) ionization source, fast atomic impact (“FAB”) ionization source, liquid secondary ion mass analysis (“LSIMS”) ionization source, desorption electrospray ionization (“DESI”” ) Ionization source, nickel 63 radiation ionization source, thermospray ionization source, atmospheric sampling glow discharge ionization (“ASGDI”) ionization source, glow discharge (“GD”) ionization source, impact ionization source, real-time direct analysis (“DART”) Ionization source, laser spray ionization (“LSI”) source, sonic spray ionization (“SSI”) source, matrix-assisted inlet ionization (“MAII”) source, solvent-assisted inlet ionization (“SAII”) source, desorption electrospray ionization Select from the group consisting of (“DESI”) sources, desorption electrocurrent focused ionization (“DEFI”) sources, laser ablation electrospray ionization (“LAESI”) sources, and surface-assisted laser desorption ionization (“SALDI”) sources. Performed by the ionized source.

The method may include capturing the sample ions in an ion trap and / or guiding the sample ions using an ion guide.

The method may include heating the collision surface.

The methods are (i) about <100 ° C., (ii) about 100-200 ° C., (iii) about 200-300 ° C., (iv) about 300-400 ° C., (v) about 400-500 ° C., (vi). About 500 to 600 ° C, (vii) about 600 to 700 ° C, (viii) about 700 to 800 ° C, (ix) about 800 to 900 ° C, (x) about 900 to 1000 ° C, (xi) about 1000 to 1100 ° C. , And (xii) may include heating the collision surface to a temperature selected from the group consisting of about> 1100 ° C.

The method may include feeding the matrix through a matrix conduit to the sample and analyzing the ions of the sample using an ion analyzer located downstream of the outlet of the matrix conduit. ..

The distance x between the outlet of the matrix conduit and the inlet of the ion analyzer is (i) about 0.1 to 0.5 mm, (ii) about 0.5 to 1.0 mm, (iii). About 1.0 to 1.5 mm, (iv) about 1.5 to 2.0 mm, (v) about 2.0 to 2.5 mm, (vi) about 2.5 to 3.0 mm, (vi) about 3 .0 to 3.5 mm, (viii) about 3.5 to 4.0 mm, (ix) about 4.0 to 4.5 mm, (x) about 4.5 to 5.0 mm, (xi) about 5.0 ~ 5.5 mm, (xii) about 5.5-6.0 mm, (xiii) about ≤6 mm, (xiv) about ≤5.5 mm, (xv) about ≤5 mm, (xvi) about ≤4.5 mm, ( It may be selected from the group consisting of (xvii) about ≦ 4 mm, (xvii) about ≦ 3.5 mm, and (xix) about ≦ 3 mm.

The distance x is about 6.0 to 6.5 mm, about 6.5 to 7.0 mm, 7.0 to 7.5 mm, about 7.5 to 8.0 mm, about 8.0 to 8.5 mm, and about 8. .5-9.0 mm, about 9.0-9.5 mm, about 9.5-10.0 mm, about 0.1-10 mm, about 0.1-7.5 mm, about 0.1-5.1 mm, It is contemplated that it may be selected from the group consisting of about 0.5-5.1 mm and about 0.5-5.0 mm.

The ion analyzer may include a vacuum chamber in which the inlet opens. The inlet of the ion analyzer may be determined to be the region of pressure in the vacuum chamber. For example, if the inlet is provided by an elongated tube, the distance x may be determined from the position within the tube, which is the pressure of the vacuum chamber. Alternatively or additionally, the inlet may be determined to be the inlet and / or outlet of the inflow pipe or orifice.

The matrix conduit may have an outlet opening substantially facing the inlet of the ion analyzer or on the opposite side of the inlet of the ion analyzer, and / or the outlet opening of the matrix conduit may be. It may be substantially coaxial with the inlet of the ion analyzer.

The outlet of the matrix conduit and the inlet of the ion analyzer need not be separated and connected by a completely enclosed tube.

The outlet of the matrix conduit and the inlet of the ion analyzer may be interconnected by a sampling tube with an opening around it to receive the sample through the opening.

The method may include delivering the sample through a sample transfer tube, the sample being placed so as to collide with the upstream side of the sampling tube, then flowing around the outside of the sampling tube and within the downstream side of the sampling tube. Enter the opening.

The outlet of the sample transfer tube may be separated from the outlet of the matrix conduit and / or the inlet of the ion analyzer and / or sampling tube and not connected to these elements by a enclosed tube.

The longitudinal axis of the sample transfer tube is substantially orthogonal to the longitudinal axis through the outlet of the matrix conduit and / or the longitudinal axis through the inlet of the ion analyzer and / or the longitudinal axis of the sampling tube. You may.

The matrix is introduced into the specimen at a distance y upstream of the inlet of the ion analyzer, or upstream of the vacuum chamber in which the collision surface is located, where y is (i) 1.5- 2.0 mm, (ii) about 2.0 to 2.5 mm, (iii) about 2.5 to 3.0 mm, (iv) about 3.0 to 3.5 mm, (v) about 3.5 to 4. 0 mm, (vi) about 4.0-4.5 mm, (vii) about 4.5-5.0 mm, (viii) about 5.0-5.5 mm, (ix) about 5.5-6.0 mm, (X) about ≧ 6 mm, (xi) about ≧ 7 mm, (xii) about ≧ 8 mm, (xiii) about ≧ 9 mm, (xiv) about ≧ 10 mm, (xv) about ≧ 12 mm, (xvi) about ≧ 14 mm, It is selected from the group consisting of (xvii) about ≧ 16 mm, (xvii) about ≧ 18 mm, (xix) about ≧ 20 mm, (xx) about ≧ 25 mm, and (xxi) about ≧ 30 mm.

The sample may be fed through the sample transfer tube, the matrix may be fed directly into the sample transfer tube, or the matrix may be fed through the matrix conduit and the sample may be fed into the matrix conduit. It may be supplied directly.

The sample transfer tube and / or matrix conduit may be directly connected to the inlet of the ion analyzer, or a vacuum chamber in which the collision surface is located.

Alternatively, the sample may be fed through the sample transfer tube and the matrix is fed to the sample downstream of the outlet of the sample transfer tube.

For example, sample movement may be provided to perform the step of providing the sample, and the outlet of the matrix conduit may be provided at a location around the sample transfer tube. The gas stream may be arranged so as to enter the sample from the outlet and further clear the matrix to the inlet of the ion analyzer that analyzes the ions.

The matrix is a matrix conduit having an inner diameter selected from the group consisting of (i) about ≤900 μm, (ii) about ≤800 μm, (iii) about ≤700 μm, (iv) about ≤600 μm, and (v) about ≤500 μm. It may be supplied through.

Alternatively, the matrix has (i) about ≤450 μm, (ii) about ≤400 μm, (iii) about ≤350 μm, (iv) about ≤300 μm, (v) about ≤250 μm, (vi) about ≤200 μm, (vi). It may be supplied through a matrix conduit having an inner diameter selected from the group consisting of (vi) about ≤150 μm, (viii) about ≤100 μm, (ix) about ≤50 μm, and (x) about ≤25 μm.

Matrix conduits with smaller inner diameters tend to produce better and less noise spectra. However, matrix ducts with inner diameters of ≧ 450 μm, ≧ 500 μm, or ≧ 1 mm are contemplated.

The matrix may be supplied through the matrix conduit and the outlet end of the matrix duct may be tapered to narrow in the downstream direction.

The matrix was 5 to 50 μl / min, (i) about 50 to 100 μl / min, (ii) about 100 to 150 μl / min, (iii) about 150 to 200 μl / min, (iv) about 200 to 250 μl / min, (i). v) Approximately 250-300 μl / min, (vi) Approximately 300-350 μl / min, (vii) Approximately 350-400 μl / min, (viii) Approximately 400-450 μl / min, (ix) Approximately 450-500 μl / min, ( x) about 500 to 550 μl / min, (xi) about 550 to 600 μl / min, (xii) about 600 to 650 μl / min, (xiii) about 650 to 700 μl / min, (xiv) 700 to 750 μl / min, (xv) ) Approximately 750 to 800 μl / min, (xvi) approximately 800 to 850 μl / min, (xvii) approximately 850 to 900 μl / min, (xviii) approximately 900 to 950 μl / min, (xix) approximately 950 to 1000 μl / min, (xx) ) At a flow rate selected from the group consisting of about 50 μl / min to 1 ml / min, (xxii) about 100 to 800 μl / min, (xxxii) about 150 to 600 μl / min, and (xxxii) about 200 to 400 μl / min. It may be supplied to the sample by a matrix conduit.

Relatively low matrix flow rates may be used so that the matrix is non-toxic and / or does not contaminate the instrument.

The method may include generating sample ions and analyzing the sample ions.

The steps of analyzing the sample ion are (i) mass spectrometrically analyzing the sample ion, (ii) analyzing the ion mobility or different ion mobility of the sample ion, and (iii) the ion cross section of the sample ion. Or to analyze the collision cross section, (iv) separate the sample ions according to their ion mobility or different ion mobility, (v) before mass spectrometrically analyzing the sample ions, remove the sample ions from their ions. Separation according to mobility or different ion mobility, or (vi) exclusion or disposal of sample ions based on their ion mobility or different ion mobility may be included.

The method is to analyze the sample ions with an ion analyzer to obtain sample ion data, to analyze rock mass, lock mobility, or calibration ions, and to calibrate the ion analyzer, or the lock. It may include adjusting the sample ion data based on the data obtained from analyzing mass, lock mobility, or calibration ions.

The rock mass, lock mobility, or calibration compound may be in the matrix to generate rock mass, lock mobility, or calibration ions.

The rock mass, lock mobility, or calibration compound / ion may be introduced into the matrix conduit, sample conduit, or supplied in a separate tube.

The method may include using a first device to provide the sample, the first device may include or form a portion of ambient ions or ionization sources, or said. The first device may generate the aerosol, smoke or vapor from the target being analyzed, contains ions and / or is subsequently ionized by ambient or ionization sources or other ionization sources.

The target may comprise a natural or unmodified target material.

Natural or unmodified targets may be unmodified (ie, unmodified) by the addition of matrices or reagents.

The first device may be arranged and adapted to generate an aerosol, smoke, or vapor from one or more regions of the target without the need for the target prior to formulation.

The first device is (i) rapid evaporation ionization mass analysis (“REIMS”) ion source, (ii) desorption electrospray ionization (“DESI”) ionization source, (iii) laser desorption ionization (“LDI”). Ion source, (iv) thermal desorption ion source, (v) laser diode thermal desorption (“LDTD”) ion source, (vi) desorption electrospray focused (“DEFI”) ion source, (vii) dielectric barrier Discharge ("DBD") plasma ion source, (viii) atmospheric solid-state analysis probe ("ASAP") ion source, (ix) ultrasonic-assisted spray ionization ion source, (x) simple atmospheric sonic spray ionization ("EASI") Ion Source, (xi) Desorption Atmospheric Pressure Photoionization (“DAPPI”) Ion Source, (xii) Paper Spray (“PS”) Ion Source, (xiii) Jet Desorption Ionization (“JeDI”) Ion Source, (xiv) ) Touch spray ("TS") ion source, (xv) nanoDISI ion source, (xvi) laser ablation electrospray ("LAESI") ion source, (xvii) real-time direct analysis ("DART") ionization source, (xviii) ) Probe Electrospray Ionization ("PESI") Ion Source, (xx) Solid Probe Assisted Electrospray Ionization ("SPA-ESI") Ion Source, (xx) Cavitron Ultrasonic Surgical Aspiration ("CUSA") Device, (xxi ) Hybrid CUSA diathermy device, (xxiii) focused or unfocused electrospray ionization device, (xxiii) hybrid focused or unfocused ultrasonic excision and diathermy device, (xxiv) microwave resonator, (xxv) pulse plasma RF dissociation device, (Xxxvi) Argon Plasma Coagulator, (xxxvi) Hybrid Pulse Plasma RF Dissociation and Argon Plasma Coagulator, (xxxvi) Hybrid Pulse Plasma RF Dissociation and JeDI Instrument, (xxxvii) Surgical Water / Physiological Salt Water Jet Device, (xxx) Hybrid A part of a device selected from the group consisting of an electrospray and argon plasma coagulation device and a (xxx) hybrid argon plasma coagulation and water / physiological saline jet device, or an ion source may be provided or formed. ..

The step of using the first device to generate an aerosol, smoke, or vapor from one or more regions of the target may include contacting the target with one or more electrodes.

The one or more electrodes may further include (i) the method providing an arbitrarily separate counter electrode, (ii) a bipolar device, or (iii) the method being arbitrarily separate. It may be equipped with any of the polyphase RF devices, further comprising providing one or more counter electrodes.

One or more electrodes may comprise or form part of a rapid evaporation ionization mass spectrometry (“REIMS”) device.

The method may include applying an AC or DC voltage to the one or more electrodes to generate the aerosol, smoke, or vapor.

The step of applying the AC or DC voltage to the one or more electrodes may include applying one or more pulses of the AC or DC voltage to the one or more electrodes.

Heat may be dissipated into the target by the step of applying the AC or DC voltage to the one or more electrodes.

The step of using the first device to generate an aerosol, smoke, or vapor from one or more regions of the target may include irradiating the target with a laser.

The first device may be arranged and adapted to generate an aerosol from one or more regions of the target by directly evaporating or vaporizing the target material from the target by Joule heat or diathermy.

Diathermy refers to three techniques: ultrasonic (ultrasonic) diathermy, radiofrequency diathermy (eg, shortwave, eg, radio frequencies in the range of 1-100 MHz), or microwave diathermy (eg, 915 MHz or 2). It may be generated by one of (in the .45 GHz band). Diathermy's methods differ primarily in their ability to penetrate.

The step of using the first device to generate an aerosol, smoke, or vapor from one or more regions of the target may include directing ultrasonic energy to the target.

Aerosols may comprise uncharged aqueous droplets and optionally include cellular material.

At least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% of the mass or substance generated by the first device and forming the aerosol. , May be in the form of droplets.

The first device may be arranged and adapted to generate an aerosol, the apparent diameter of the aerosol (“SMD”, d32) being (i) <5 μm, (ii) 5-10 μm, (i). It is in the range of iii) 10 to 15 μm, (iv) 15 to 20 μm, (v) 20 to 25 μm, or (vi)> 25 μm.

Aerosols have Reynolds numbers (Re) in the range of (i) <2000, (ii) 2000-2500, (iii) 2500-3000, (iv) 3000-3500, (v) 3000-4000, or (vi)> 4000. ) May be transferred.

In terms of substantially generating the aerosol, the aerosol is (i) <50, (ii) 50-100, (iii) 100-150, (iv) 150-200, (v) 200-250, (v) vi) 250-300, (vii) 300-350, (viii) 350-400, (ix) 400-450, (x) 450-500, (xi) 500-550, (xii) 550-600, (xiii) ) 600-650, (xiv) 650-700, (xv) 700-750, (xvi) 750-800, (xvi) 800-850, (xvii) 850-900, (xix) 900-950, (xx) It may include droplets having a Weber number (We) selected from the group consisting of 950-1000 and (xxi)> 1000.

In terms of substantially generating the aerosol, the aerosol is (i) 1-5, (ii) 5-10, (iii) 10-15, (iv) 15-20, (v) 20-25, (vi) 25~30, (vii) 30~35, (viii) 35~40, (ix) 40~45, (x) 45~50, and (xi)> 50 Stokes number in the range of _(S k) May include droplets having.

In terms of substantially generating the aerosol, the aerosol is (i) <20 m / s, (ii) 20-30 m / s, (iii) 30-40 m / s, (iv) 40-50 m / s, (V) 50-60 m / s, (vi) 60-70 m / s, (vii) 70-80 m / s, (viii) 80-90 m / s, (ix) 90-100 m / s, (x) 100- It consists of 110 m / s, (xi) 110-120 m / s, (xii) 120-130 m / s, (xiii) 130-140 m / s, (xiv) 140-150 m / s, and (xv)> 150 m / s. It may comprise droplets having an mean axial flow velocity selected from the group.

The target may include biological tissue such as healthy or diseased tissue.

Biological tissue may comprise human tissue or non-human animal tissue.

The biological tissue may include a biological tissue in the body.

The biological tissue may include an extracorporeal biological tissue.

The biological tissue may include in vitro biological tissue.

Biological tissues include adrenal tissue, worm tissue, bladder tissue, bone, intestinal tissue, brain tissue, breast tissue, bronchi, coronary tissue, ear tissue, esophageal tissue, eye tissue, bile sac tissue, genital tissue, heart tissue, hypothalamic tissue. , Kidney tissue, colon tissue, intestinal tissue, pharyngeal tissue, liver tissue, lung tissue, lymph node, oral tissue, nasal tissue, pancreatic tissue, parathyroid tissue, pituitary tissue, prostate tissue, rectal tissue, salivary gland tissue, skeletal muscle Tissue, skin tissue, small intestine tissue, spinal tissue, spleen tissue, gastric tissue, thoracic gland tissue, tracheal tissue, thyroid tissue, soft tissue, connective tissue, peritoneal tissue, vascular tissue, adipose tissue, urinary tract tissue, urinary tract tissue, stage I, It may include stage II, stage III or stage IV cancer tissue, metastatic cancer tissue, mixed stage cancer tissue, substage cancer tissue, healthy or normal tissue, or cancer or abnormal tissue.

The first device may include a point of care (“POC”), diagnostic, or surgical device.

The method may include ionizing at least a portion of the aerosol, smoke, or vapor to generate sample ions.

The method may include directing or aspirating at least a portion of the aerosol, smoke, or vapor to the vacuum chamber of the mass and / or ion mobility analyzer.

The method may also include ionizing one of the mass and / or ion mobility analyzers or at least a portion of the aerosol, smoke, or vapor in the vacuum chamber to generate multiple sample ions. Good.

The method comprises arbitrarily colliding the aerosol, smoke, or vapor with a collision surface placed in the vacuum chamber of the mass and / or ion mobility analyzer to generate multiple sample ions. It may be.

The method may include mass and / or ion mobility analysis of the sample ions or ions drawn from the aerosol, smoke, or vapor in order to obtain mass and / or ion mobility analysis data.

The methods are (i) to distinguish between healthy and lesioned tissue, (ii) to distinguish between potential cancerous tissue and non-cancerous tissue, and (iii) to distinguish between different types or degrees of cancerous tissue. (Iv) To distinguish between different types or classes of target material, (v) to determine if one or more desirable or undesired substances are present within the target, (vi) to determine the presence of the target. To confirm identification or reliability, (vii) to determine if one or more impurities, illegal substances, or unwanted substances are present within the target, (vii) human or animal patients To determine if the risk of adverse outcomes has increased, (ix) to make a diagnosis or future prediction, or to assist in making, and (x) a surgeon, nurse, doctor, or It may include analyzing the analytical data for any purpose of notifying the robot of medical, surgical, or diagnostic results.

The method may be either a surgical method or part of a non-surgical method. For example, the method may be a surgical method in which the sample may be human or animal tissue containing the sample. The sample may undergo electrosurgical diathermy evaporation, or other form of rapid evaporation, to form a gas phase sample, vapor sample, or aerosol. As an example, devices and methods may be used to identify human cells in breast cancer surgery. By analyzing the sample ions, it is possible to determine whether or not the tissue is cancerous.

Alternatively, the method may include a non-surgical method. For example, human or animal tissue that is not part of the human or animal body (ie, pre-extracted, placed, or removed) may be analyzed, or a sample or biological tissue other than human or animal tissue is analyzed. You may. Also, by analyzing the sample ions, it is possible to determine the characteristics or components of the sample, such as whether or not the sample contains cancer tissue.

Embodiments include country of origin identification, drug testing, food safety testing (eg dairy products), cosmetic testing, military use, air pollution testing, post-mortem analysis, microbial identification (eg bacteria), and other non-surgical embodiments such as automated sampling. It may be used in the method.

The method may be used to analyze abiotic samples and compounds. The sample formed from the sample may be partially charged and / or may have a relatively high organic content.

The step of analyzing the analytical data may include analyzing one or more sample spectra, such as classifying aerosol, smoke, or vapor samples.

Analyzing one or more sample spectra, such as classifying aerosol, smoke, or vapor samples, is an unsupervised analysis of one or more sample spectra (eg, degeneracy), and / or 1 It may include an analysis (eg, classification) of one or more sample spectra with objective variables.

Analyzing one or more sample spectra may include analysis without objective variable (eg, degeneracy) followed by analysis with objective variable (eg, classification).

Analyzing one or more sample spectra includes (i) univariate analysis, (ii) multivariate analysis, (iii) principal component analysis (PCA), (iv) linear discriminant analysis (LDA), (v). Principal component criteria (MMC), (vi) library-based analysis, (vii) soft-less modeling for classification (SIMCA), (viii) factor analysis (FA), (ix) recursive division (decision tree), (X) Random Forest, (xi) Independent Principal Component Analysis (ICA), (xii) Partriminant Squared Discriminant Analysis (PLS-DA), (xiii) Linear (Partial Minimal Squared) Projection (OPLS), (xiv) OPLS Discriminant Analysis (OPLS-DA), (xv) Support Vector Machine (SVM), (xvi) (Artificial) Neural Network, (xvii) Multilayer Perceptron, (xvii) Radiation Base Function (RBF) Network, (xix) Bayes Analysis, (xx) cluster analysis, (xxii) kernel method, (xxxii) partial discriminant analysis, (xxiii) k neighborhood method (KNN), (xxiv) secondary discriminant analysis (QDA), (xxv) established principal component analysis Use one or more of (PPCA), (xxvi) non-negative matrix factor decomposition, (xxvii) k mean factor decomposition, (xxxviii) fuzzy c mean factor decomposition, and (xxx) discriminant analysis (DA). May include.

Analyzing one or more sample spectra to classify aerosol, smoke, or vapor samples may include developing a classification model or library using one or more standard sample spectra. ..

Analyzing one or more sample spectra to classify aerosol, smoke, or steam samples is performed after performing principal component analysis (PCA) (eg, for classification). It may include performing a linear discriminant analysis (LDA).

Analyzing one or more sample spectra to classify aerosol, smoke, or vapor samples is performed after performing Principal Component Analysis (PCA) (eg, for classification). It may include performing a maximum margin criterion (MMC).

Analyzing one or more sample spectra to classify aerosol, smoke, or vapor samples may include defining one or more classes within the classification model or library.

Analyzing one or more sample spectra to classify aerosol, smoke, or vapor samples can be done with one or more classes in a classification model or library according to one or more class or cluster criteria. It may include demarcating manually or automatically.

One or more class or cluster criteria for each class is the distance between one or more pairs of reference points with respect to the standard sample spectrum in model space, within a group of reference points with respect to the standard sample spectrum in model space. It may be based on the variance value, as well as one or more of the variance values in the group of reference points relative to the standard sample spectrum in model space.

One or more classes may each be defined by one or more class definitions.

One or more class definitions are a set of one or more reference points for standard sample spectra, values, boundaries, lines, planes, hyperplanes, variances, volumes, Voronoi cells, and / or positions in model space. , And one or more of one or more positions in the class hierarchy.

Analyzing one or more sample spectra to classify aerosol, smoke, or vapor samples involves using a classification model or library to classify one or more unknown sample spectra. It may be.

Analyzing one or more sample spectra to classify aerosol, smoke, or vapor samples manually or automatically classifies one or more sample spectra according to one or more classification criteria. May include doing.

One or more classification criteria
One or more projected sample points for one or more sample spectra in model space, and standard sample spectra, values, boundaries, lines in model space that are below or at such a minimum distance. Distance between a plane, hyperplane, volume, Voronoi cell, or a set of one or more reference points relative to a position,
One or more standard sample spectra in model space, one or more sides of one or more reference points relative to a value, boundary, line, plane, hyperplane, or position in model space. Position with respect to one or more projected sample points for one or more sample spectra,
Position for one or more projected sample points with respect to one or more volume volumes in the model space or one or more sample spectra in the model space within the Voronoi cell,
It may include one or more of the probabilities or classification scores that are higher than the probability or classification score threshold, or that are such highest probabilities or classification scores.

Mass and / or ion mobility analysis may acquire data in negative ion mode only, in positive ion mode only, or in both positive and negative ion modes. The positive ion mode analysis data may be combined with or concatenated with the negative ion mode analysis data. Negative ion mode can provide a particularly useful spectrum for classifying aerosol, smoke, or vapor samples, such as aerosol, smoke, or vapor samples from lipid-laden targets.

Ionic conductivity analysis data may be obtained using drift gases of different ion mobility and / or dopants are added to the drift gas to induce a change in drift time of one or more species. You may. This data may then be combined or concatenated.

The first aspect of the present invention is
The sample inlet for receiving the sample and
A matrix inlet for receiving the matrix compound and
A mixing region for mixing the sample with the matrix compound, such that the sample is diluted by the matrix and dissolved in the matrix during use, or forms a first cluster in the matrix.
Collision surface and
Perform mass and / or ion mobility analysis with the first cluster or first droplet of diluted or lysed specimen with a device adapted and arranged to collide with the collision surface. Equipment for doing so is also provided.

The device may be arranged and adapted to cause the first cluster or first droplet to collide with the collision surface and break into a plurality of second smaller clusters or droplets.

The device may be configured to provide the sample in the form of a gas phase sample, aerosol, vapor, smoke, or lipid and / or supply the matrix to the sample, wherein the sample is a gas phase sample, aerosol. , Vapor, smoke, or lipid form.

The device may be configured to feed the matrix in the form of an aerosol, vapor or smoke, while the matrix is in the form of an aerosol, vapor or solid particles.

The device may be configured to feed the matrix molecules to the sample and intermix the matrix molecules with the sample, the matrix being in the gas phase or of aerosol, vapor, or solid particles. It is a shape.

The device may include a high pressure region and a low pressure region, the gas phase matrix cools in use, condenses into a liquid, and dissolves in the droplet matrix such that the sample forms the first droplet. As such, the mixture of the sample and the matrix may be configured to be transferred from the high pressure region to the constant pressure region.

Alternatively, the device may be configured to feed the matrix as a liquid sample and intermix with the sample.

If the specimen and / or matrix is in the form of a liquid, the device may be configured to convert the specimen and / or matrix into droplets or vapors, for example by jetting or spraying. For example, if the sample and matrix are mixed as a liquid, the mixture may be subsequently converted into a first sample droplet, for example by spraying or spraying.

The matrix is first fed as a solid (eg powder), sublimated or melted and evaporated to form a matrix in a vapor or gas phase that is intermixed with the sample to form clusters of the sample and matrix. You may.

The matrix comprises (i) a solvent for the sample, (ii) an organic solvent, (iii) a volatile compound, (iv) a polar molecule, (v) water, (vi) one or more alcohols, (vii). Methanol, (viii) ethanol, (ix) isopranol, (x) acetone, (xi) acetonitrile, (xii) 1-butanol, (xiii) tetrahydrofuran, (xiv) ethyl acetate, (xv) ethylene glycol, (xvi) It may be selected from the group consisting of dimethyl sulfoxide, (xvii) aldehyde, (xvii) ketone, (xiv) non-polar molecule, (xx) hexane, (xxx) chloroform, (xxxi) butanol, and (xxiii) propanol. ..

By way of example, for specimens with polar lipids, the low molecular weight alcohol may be used as a matrix (eg methanol, ethanol, isopropanol) or a ketone (eg acetone). These matrices have been shown to enhance species ionization, which would otherwise be detected as low intensity and no vapor in the matrix.

The solvent of the proton matrix may be used, for example, for the analysis of lipids or triglycerides. Alternatively, aprotonic or aproton matrix solvents may be used, for example, for protein analysis.

The mixture of sample and matrix may be a homogeneous mixture or a heterogeneous mixture.

The matrix and / or the sample may be doped with one or more additives to increase the solventization of the sample in the matrix or to increase the ionization of the sample.

The matrix or specimen is doped with acidic or basic additives. For example, the matrix may be doped with formic acid, diethylamine.

The matrix may result in derivatization of the specimen. For example, the matrix may result in the derivatization of cholesterol or steroids within the specimen. Thereby, the ionized sample may be given more easily.

The calibrator may be added to the matrix and / or specimen, and the compounds in the matrix and / or specimen may be selected as calibrators to calibrate the method of mass spectrometry and / or ion mobility analysis. May be used for. This is particularly beneficial for ambient ionization techniques such as the REIMS technique, which can make it difficult to introduce the calibrator according to conventional techniques.

The device may be configured to accelerate the first cluster or first droplet to the collision surface.

The device may be configured to use the pressure difference to accelerate the first cluster or first droplet to the collision surface.

The device is configured to create a pressure difference between the two regions in order to accelerate the first cluster or first droplet between the first region and the second region to the collision surface. You may.

The device may include a mass and / or ion mobility analyzer with an atmospheric interface located between the first and second regions, the second region being connected to a vacuum pump. A vacuum chamber for accommodating the collision surface may be provided.

The device may be configured to evaporate the matrix from the sample in the second droplet so as to provide the sample ions separated from the matrix.

The step of colliding the first cluster or the first droplet with the collision surface may evaporate the matrix from the sample by converting the kinetic energy of the sample and matrix into heat.

The step of colliding the first cluster or first droplet may produce a second smaller cluster or droplet, at least a portion of which may have only a single sample molecule. This improves the ionization process.

The step of evaporating the matrix from the sample may be charge transfer to or from the sample such that the sample is ionized to form sample ions.

The device may include sample ions to guide the sample ions and / or ion traps to capture the ion guides.

The device may be equipped with a heater to heat the collision surface.

The heaters are (i) about <100 ° C, (ii) about 100-200 ° C, (iii) about 200-300 ° C, (iv) about 300-400 ° C, (v) about 400-500 ° C, (vi). ) About 500 to 600 ° C., (vii) about 600 to 700 ° C., (viii) about 700 to 800 ° C., (ix) about 800 to 900 ° C., (x) about 900 to 1000 ° C., (xi) about 1000 to 1100. It may be configured to heat the collision surface to a temperature selected from the group consisting of ° C. and (xi) about> 1100 ° C.

The device may include a matrix conduit for supplying the matrix to the sample and an ion analyzer located downstream of the outlet of the matrix conduit for analyzing the ions of the sample.

The distance x between the outlet of the matrix conduit and the inlet of the ion analyzer is (i) about 0.1 to 0.5 mm, (ii) about 0.5 to 1.0 mm, (iii). About 1.0 to 1.5 mm, (iv) about 1.5 to 2.0 mm, (v) about 2.0 to 2.5 mm, (vi) about 2.5 to 3.0 mm, (vi) about 3 .0 to 3.5 mm, (viii) about 3.5 to 4.0 mm, (ix) about 4.0 to 4.5 mm, (x) about 4.5 to 5.0 mm, (xi) about 5.0 ~ 5.5 mm, (xii) about 5.5-6.0 mm, (xiii) about ≦ 6 mm, (xiv) about 5.5 ≦ mm, (xv) about 5 ≦ mm, (xvi) about 4.5 ≦ It may be selected from the group consisting of mm, (xvii) about 4 ≦ mm, (xvii) about 3.5 ≦ mm, and (xix) about 3 ≦ mm.

The ion analyzer may include a vacuum chamber in which the inlet opens. The inlet of the ion analyzer may be determined to be the region of pressure in the vacuum chamber. For example, if the inlet is provided by an elongated tube, the distance x may be determined from the position within the tube, which is the pressure of the vacuum chamber. Alternatively or additionally, the inlet may be determined to be the inlet and / or outlet of the inflow pipe or orifice.

The matrix conduit may have an outlet opening substantially facing the inlet of the ion analyzer or on the opposite side of the inlet of the ion analyzer, and / or the outlet opening of the matrix conduit may be. It is substantially coaxial with the inlet of the ion analyzer.

The outlet of the matrix conduit and the inlet of the ion analyzer need not be separated and connected by a completely enclosed tube.

The outlet of the matrix conduit and the inlet of the ion analyzer may be interconnected by a sampling tube with an opening around it to receive the sample through the opening.

The device may be equipped with a sample transfer tube for delivering the sample, in which the sample crashes into the upstream side of the sampling tube during use and flows around the outside of the sampling tube, on the downstream side of the sampling tube. It may be arranged so as to enter the opening.

The outlet of the sample transfer tube may be separated from the outlet of the matrix conduit and / or the inlet of the ion analyzer and / or sampling tube and not connected to these elements by a enclosed tube.

The longitudinal axis of the sample transfer tube is substantially orthogonal to the longitudinal axis through the outlet of the matrix conduit and / or the longitudinal axis through the inlet of the ion analyzer and / or the longitudinal axis of the sampling tube. You may.

The matrix may be introduced into the specimen at a distance y upstream of the inlet of the ion analyzer, or upstream of the vacuum chamber in which the collision surface is located, where y is (i) 1. .5-2.0 mm, (ii) about 2.0-2.5 mm, (iii) about 2.5-3.0 mm, (iv) about 3.0-3.5 mm, (v) about 3.5 ~ 4.0 mm, (vi) about 4.0-4.5 mm, (vii) about 4.5-5.0 mm, (viii) about 5.0-5.5 mm, (ix) about 5.5-6 .0 mm, (x) about ≧ 6 mm, (xi) about ≧ 7 mm, (xii) about ≧ 8 mm, (xiii) about ≧ 9 mm, (xiv) about ≧ 10 mm, (xv) about ≧ 12 mm, (xvi) about ≧ It is selected from the group consisting of 14 mm, (xvii) about ≧ 16 mm, (xvii) about ≧ 18 mm, (xix) about ≧ 20 mm, (xx) about ≧ 25 mm, and (xxi) about ≧ 30 mm.

The device may include a sample transfer tube for supplying the sample and a matrix conduit for supplying the matrix directly into the sample transfer tube, or a matrix conduit for supplying the matrix, and a matrix conduit for the sample. A sample transfer tube for direct supply may be provided therein.

The sample transfer tube and / or matrix conduit may be directly connected to the inlet of the ion analyzer, or a vacuum chamber in which the collision surface is located.

Alternatively, the sample may be fed through the sample transfer tube and the matrix is fed to the sample downstream of the outlet of the sample transfer tube. For example, a sample transfer tube may be provided to perform the step of providing the sample, and the outlet of the matrix conduit may be provided at a location around the sample transfer tube. The gas stream may be arranged so as to clear the matrix from the outlet into the sample and at the inlet of the ion analyzer that analyzes the ions.

The device may include a matrix conduit for supplying the matrix, which are (i) about ≤450 μm, (ii) about ≤400 μm, (iii) about ≤350 μm, (iv) about ≤300 μm, (i). v) An inner diameter selected from the group consisting of about ≤250 μm, (vi) about ≤200 μm, (vi) about ≤150 μm, (viii) about ≤100 μm, (ix) about ≤50 μm, and (x) about ≤25 μm. Have.

Matrix conduits with smaller inner diameters tend to produce better and less noise spectra.

The device may include a matrix conduit for supplying the matrix, and the outlet end of the matrix conduit is tapered to narrow in the downstream direction.

The apparatus is 5 to 50 μl / min, (i) about 50 to 100 μl / min, (ii) about 100 to 150 μl / min, (iii) about 150 to 200 μl / min, (iv) about 200 to 250 μl / min, (i). v) About 250-300 μl / min, (vi) about 300-350 μl / min, (vii) about 350-400 μl / min, (viii) about 400-450 μl / min, (ix) about 450-500 μl / min, (ix) x) about 500 to 550 μl / min, (xi) about 550 to 600 μl / min, (xii) about 600 to 650 μl / min, (xiii) about 650 to 700 μl / min, (xiv) 700 to 750 μl / min, (xv) ) Approximately 750 to 800 μl / min, (xvi) approximately 800 to 850 μl / min, (xvii) approximately 850 to 900 μl / min, (xviii) approximately 900 to 950 μl / min, (xix) approximately 950 to 1000 μl / min, (xx) ) Matrix at a flow rate selected from the group consisting of about 50 μl / min to 1 ml / min, (xxii) about 100 to 800 μl / min, (xxxii) about 150 to 600 μl / min, and (xxxii) about 200 to 400 μl / min. A pump configured to feed the matrix through a conduit may be provided.

Relatively low matrix flow rates may be used so that the matrix is non-toxic and / or does not contaminate the instrument.

The device may include a mass and / or ion mobility analyzer for analyzing sample ions.

The device may include a first device for providing the specimen, the first device comprising or forming a portion of ambient ions or ionization sources, or the first device. It is configured to produce one or the aerosol, smoke, or vapor from the target being analyzed, contains ions, and / or is subsequently ionized by ambient or ionization sources or other ionization sources.

The target may comprise a natural or unmodified target material.

Natural or unmodified target materials are not modified by the addition of matrices or reagents.

The first device or device is arranged and adapted to generate an aerosol, smoke, or vapor from one or more regions of the target without the need for the target prior to formulation.

The first device or apparatus is (i) rapid evaporation ionization mass analysis (“REIMS”) ion source, (ii) desorption electrospray ionization (“DESI”) ionization source, (iii) laser desorption ionization (“LDI”). ”) Ion source, (iv) thermal desorption ion source, (v) laser diode thermal desorption (“LDTD”) ion source, (vi) desorption electrospray focused (“DEFI”) ion source, (vii) dielectric Body Barrier Discharge (“DBD”) Plasma Ion Source, (viii) Atmospheric Solid Analytical Probe (“ASAP”) Ion Source, (ix) Ultrasonic Assisted Spray Ionization Ion Source, (x) Simple Atmospheric Sonic Spray Ionization (“EASI”) ”) Ion source, (xi) desorption atmospheric pressure photoionization (“DAPPI”) ion source, (xii) paper spray (“PS”) ion source, (xiii) jet desorption ionization (“JeDI”) ion source, (Xiv) Touch Spray ("TS") Ion Source, (xv) Nano DISI Ion Source, (xvi) Laser Ablation Electrospray ("LAESI") Ion Source, (xvi) Real Time Direct Analysis ("DART") Ion Source, (Xviii) Probe Electrospray Ionization (“PESI”) Ion Source, (xx) Solid Probe Assisted Electrospray Ionization (“SPA-ESI”) Ion Source, (xx) Cavitron Ultrasonic Surgical Aspiration (“CUSA”) Device, (Xxi) Hybrid CUSA diathermy device, (xxiii) focused or unfocused electrospray ionization device, (xxxii) hybrid focused or unfocused electrospray and diathermy device, (xxiv) microwave resonance device, (xxv) pulse plasma RF dissociation Equipment, (xxxvi) argon plasma coagulation device, (xxvi) hybrid pulse plasma RF dissociation and argon plasma coagulation device, (xxvii) hybrid pulse plasma RF dissociation and JeDI device, (xxxvii) surgical water / physiological saline jet device, (xxix) ) Part of a device selected from the group consisting of a hybrid electrospray and argon plasma coagulation device, and a (xxx) hybrid argon plasma coagulation and water / physiological saline jet device, or an ion source. May be good.

The first device or device may include one or more electrodes, and by contacting the target with the one or more electrodes, aerosol, vapor, or from one or more regions of the target. It may be arranged and adapted to generate vapor.

One or more electrodes are provided with (i) an arbitrarily separate counter electrode, a unipolar device, (ii) a bipolar device, or (iii) arbitrarily at least one separate counter electrode. It may be equipped with any of the polyphase RF devices.

One or more electrodes may be equipped with a rapid evaporation ionization mass spectrometry (“REIMS”) device.

The device may include a device arranged and adapted to apply an AC or DC voltage to the one or more electrodes to generate the aerosol, smoke, or vapor.

A device for applying the AC or DC voltage to the one or more electrodes is arranged to apply one or more pulses of the AC or DC voltage to the one or more electrodes. You may.

Heat may be dissipated into the target by applying the AC or DC voltage to the one or more electrodes.

The first device or device may include a laser for irradiating the target.

The first device may be arranged and adapted to generate an aerosol from one or more regions of the target by direct evaporation or vaporization of the target material from the target by Joule heat or diathermy.

The first device or device may be arranged and adapted to direct ultrasonic energy towards the target.

Aerosols may comprise uncharged aqueous droplets and optionally include cellular material.

At least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95 of the mass or substance generated by the first device or apparatus and forming the aerosol. % May be in the form of droplets.

The first device may be arranged and adapted to generate an aerosol, the apparent diameter of the aerosol (“SMD”, d32) being (i) <5 μm, (ii) 5-10 μm, (i). It is in the range of (iii) 10 to 15 μm, (iv) 15 to 20 μm, (v) 20 to 25 μm, and (vi)> 25 μm.

Aerosols have Reynolds numbers (Re) in the range of (i) <2000, (ii) 2000-2500, (iii) 2500-3000, (iv) 3000-3500, (v) 3000-4000, or (vi)> 4000. ) May be transferred.

In terms of substantially generating the aerosol, the aerosol is (i) <50, (ii) 50-100, (iii) 100-150, (iv) 150-200, (v) 200-250, (v) vi) 250-300, (vii) 300-350, (viii) 350-400, (ix) 400-450, (x) 450-500, (xi) 500-550, (xii) 550-600, (xiii) ) 600-650, (xiv) 650-700, (xv) 700-750, (xvi) 750-800, (xvi) 800-850, (xvii) 850-900, (xix) 900-950, (xx) It may include droplets having a Weber number (We) selected from the group consisting of 950-1000 and (xxi)> 1000.

In terms of substantially generating the aerosol, the aerosol is (i) 1-5, (ii) 5-10, (iii) 10-15, (iv) 15-20, (v) 20-25, (vi) 25~30, (vii) 30~35, (viii) 35~40, (ix) 40~45, (x) 45~50, and (xi)> 50 Stokes number in the range of _(S k) May include droplets having.

In terms of substantially generating the aerosol, the aerosol is (i) <20 m / s, (ii) 20-30 m / s, (iii) 30-40 m / s, (iv) 40-50 m / s, (V) 50-60 m / s, (vi) 60-70 m / s, (vii) 70-80 m / s, (viii) 80-90 m / s, (ix) 90-100 m / s, (x) 100- It consists of 110 m / s, (xi) 110-120 m / s, (xii) 120-130 m / s, (xiii) 130-140 m / s, (xiv) 140-150 m / s, and (xv)> 150 m / s. It may comprise droplets having an mean axial flow velocity selected from the group.

The target may include biological tissue.

Biological tissue may comprise human tissue or non-human animal tissue.

The biological tissue may include a biological tissue in the body.

The biological tissue may include an extracorporeal biological tissue.

The biological tissue may include in vitro biological tissue.

Biological tissues include adrenal tissue, worm tissue, bladder tissue, bone, intestinal tissue, brain tissue, breast tissue, bronchi, coronary tissue, ear tissue, esophageal tissue, eye tissue, bile sac tissue, genital tissue, heart tissue, hypothalamic tissue. , Kidney tissue, colon tissue, intestinal tissue, pharyngeal tissue, liver tissue, lung tissue, lymph node, oral tissue, nasal tissue, pancreatic tissue, parathyroid tissue, pituitary tissue, prostate tissue, rectal tissue, salivary gland tissue, skeletal muscle Tissue, skin tissue, small intestine tissue, spinal tissue, spleen tissue, gastric tissue, thoracic gland tissue, tracheal tissue, thyroid tissue, soft tissue, connective tissue, peritoneal tissue, vascular tissue, adipose tissue, urinary tract tissue, urinary tract tissue, stage I, It may include stage II, stage III or stage IV cancer tissue, metastatic cancer tissue, mixed stage cancer tissue, substage cancer tissue, healthy or normal tissue, or cancer or abnormal tissue.

The first device or device may include a point of care (“POC”), diagnostic, or surgical device.

The collision surface may be substantially spherical, coiled, spiral, spiral, cylindrical, tubular, rod, hemispherical, teardrop-shaped, plate-shaped, concave, dish-shaped, or conical. , Or the collision surface is the inner surface of the hollow collision assembly.

The present invention provides a mass and / or ion mobility analyzer equipped with a device as described herein.

The analyzer may include a main housing or assembly of the analyzer, and the source housing may be connected to the main housing of the analyzer during use.

The analyzer may be equipped with an ion trap and / or an ion guide, which is arbitrarily configured to apply to an electric field that separates the ions from the neutral species.

The analyzer may be equipped with an analyzer for analyzing sample ions.

The analyzers are (i) a mass spectrometer for mass spectrometry of the sample ion, (ii) an ion mobility or different ion mobility analyzer, and (iii) for analyzing an ion cross section or collision cross section of the sample ion. Analyzer, (iv) Separator for separating the sample ions according to their ion mobility or different ion mobility, (v) Transfer the sample ions to them before mass spectrometry. Equipped with a separator for separation according to degree or different ion conductivity, or (vi) a device adapted to eliminate or discard sample ions based on their ion mobility or different ion mobility. May be good.

The device and analyzer may be arranged and configured to perform any one of the methods described herein.

The present invention also provides a method of surgery or electrosurgery that includes all the method steps described herein.
Bringing biological tissue into contact with surgical or electrosurgical tools and activating the tools to generate smoke, sprays, liquids, gases, surgical smoke, aerosols, or vapors containing the specimen.
Inhaling the smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor,
Mixing the matrix with the aspirated smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor.
The aspirated smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor combined with the matrix was placed in the vacuum chamber of a mass and / or ion mobility analyzer to form sample ions. Crashing into the collision surface and
Includes mass and / or ion mobility analysis of the sample ions.

The present invention is
Surgical or electrosurgical instruments containing one or more electrodes,
A device arranged and adapted to activate the device when it collides with biological tissue during use so as to generate specimen, smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor. When,
With a device adapted to aspirate the sample, smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor.
With a device adapted to mix the matrix with the aspirated specimen, smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor.
(I) A collision surface located in the vacuum chamber of the analyzer, where the specimen, smoke, spray, liquid, gas, surgical smoke, aerosol, or steam collides so as to form sample ions during use. A collision surface arranged to collide with the surface, and (ii) mass and / or ion transfer, comprising a mass and / or ion mobility analyzer for mass and / or ion mobility analysis of the sample ion. A surgical or electrosurgical device with a degree analyzer is also provided.

The surgical or electrosurgical device may be configured to perform any of the methods described herein.

Although the matrix-specimen mixture has been described above as colliding with the collision surface, it has been found that there may be improvements in sample ionization without the mixture colliding with the collision surface. The mixture may be atomized using techniques other than, for example, the mixture crashing into the collision surface. Therefore, the step of colliding the first cluster or first droplet of the diluted or dissolved matrix with the collision surface is not an essential feature of the present invention.

Therefore, from the second aspect, the present invention
Providing samples and
Supplying the sample with the matrix compound such that the sample is diluted by the matrix, dissolved in the matrix, or forms a first cluster in the matrix.
Mass and / or including breaking or collapsing the first cluster or first droplet of the diluted or dissolved matrix into a plurality of second smaller clusters or droplets. Methods for ion mobility analysis are also provided.

The first cluster or first droplet may be crushed or disintegrated by glow discharge technology or by photoionization by applying ultrasonic energy by applying laser irradiation.

The method of the second aspect may include all the features described with respect to the first aspect of the invention. However, this does not apply when the mixture does not collide with the collision surface.

For example, the method may include ionizing a sample within the second smaller cluster or droplet, or a sample drawn from the second smaller cluster or droplet. Arbitrarily, the ionization is corona discharge ionization source, reagent ionization source, photoionization source, chemical ionization source, electric shock (“EL”) ionization source, electric field ionization (“FI”) source, electric field desorption (“FD”). ”) Ionization source, inductively coupled plasma (“ICP”) ionization source, fast atomic impact (“FAB”) ionization source, liquid secondary ion mass analysis (“LSIMS”) ionization source, desorption electrospray ionization (“DESI”” ) Ionization source, nickel 63 radiation ionization source, thermospray ionization source, atmospheric sampling glow discharge ionization (“ASGDI”) ionization source, glow discharge (“GD”) ionization source, impact ionization source, real-time direct analysis (“DART”) Ionization source, laser spray ionization (“LSI”) source, sonic spray ionization (“SSI”) source, matrix-assisted inlet ionization (“MAII”) source, solvent-assisted inlet ionization (“SAII”) source, desorption electrospray ionization Select from the group consisting of (“DESI”) sources, desorption electrocurrent focused ionization (“DEFI”) sources, laser ablation electrospray ionization (“LAESI”) sources, and surface-assisted laser desorption ionization (“SALDI”) sources. Performed by the ionized source.

A second aspect of the invention is an apparatus for performing mass and / or ion mobility analysis.
A mixing region for mixing the sample with the matrix compound, such that the sample is diluted by the matrix, dissolved in the matrix, or forms a first cluster in the matrix during use.
With a device adapted to disrupt or disintegrate the first cluster or first droplet of the diluted or dissolved matrix into multiple second smaller clusters or droplets. Also provided with equipment.

The device may include a sample inlet for receiving the sample and / or a matrix inlet for receiving the matrix compound.

The device according to the second aspect may include all the features described with respect to the first aspect. Except where the device does not need to be equipped with a device adapted to collide the collision surface, or the first cluster or droplet of diluted or dissolved specimen, with the collision surface.

The present invention also provides a mass and / or ion mobility analyzer equipped with such a device.

The present invention also provides a method of surgery or electrosurgery, including the method of the second aspect.
Bringing biological tissue into contact with surgical or electrosurgical tools and activating the tools to generate smoke, sprays, liquids, gases, surgical smoke, aerosols, or vapors containing the specimen.
Inhaling the smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor,
Mixing the matrix with the aspirated smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor.
Crushing or collapsing a first cluster or droplet of a mixture to form multiple second smaller clusters or droplets.
Forming sample ions from a second cluster or droplet,
Includes mass and / or ion mobility analysis of the sample ions.

The present invention also provides a surgical or electrosurgical device comprising the device of the second aspect.
Surgical or electrosurgical instruments containing one or more electrodes,
The tool is arranged and adapted to activate the tool in the event of a collision with biological tissue during use, to generate smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor containing the specimen. Devices and
With a device adapted to aspirate the sample, smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor.
With a device adapted to mix the matrix with the sample, smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor.
With a device adapted to crush or disintegrate a first cluster or droplet of a mixture in order to form multiple second smaller clusters or droplets.
With a device arranged and adapted to ionize the specimen to form specimen ions from a second cluster or droplet.
It is equipped with a mass and / or ion mobility analyzer for analyzing the sample ions.

According to one aspect, a method of rapid evaporation ionized mass spectrometry (“REIMS”) is provided. This method provides a sample, supplies the matrix compound to the sample such that the sample dissolves in the matrix to form a first dissolved sample droplet, and the first dissolved sample. It involves colliding the first dissolved sample droplet with a collision surface or gas so as to break the sample droplet into a plurality of second smaller dissolved sample droplets.

The REIMS technique of the collision ion generator described in the Background Techniques chapter produces a sample of aqueous droplets consisting of aqueous droplets covered with electrode lipids. Aqueous droplets are accelerated by the expansion of the free jet within the atmospheric inlet of the mass spectrometer so that the fast droplets collide with the collision surface or other gas particles that generate gas ions of polar lipid molecules. However, the ionization yield of this technique is relatively low.

It has been recognized that the ion yields of conventional methods are relatively low, mainly due to the poor rate of droplet conversion to individual molecular species, mainly due to strong intermolecular bonds between sample molecules.

One embodiment dissolves the sample in a matrix, thereby substantially removing intermolecular bonds between sample molecules. Thus, when a dissolved sample subsequently collides with a collision surface or gas, as if crushing into droplets, every given droplet would contain fewer sample molecules than would have been contained in the absence of the matrix. Tends to contain.

Therefore, when the matrix in each droplet is evaporated by the method according to one embodiment, the generation of ions becomes more efficient.

The first step of colliding the dissolved sample droplets with the collision surface or gas causes the step of evaporating the matrix from the sample by converting the kinetic energy of the sample and the matrix into heat.

The first step of colliding the dissolved sample droplets may generate smaller dissolved sample droplets, in which at least a portion of the sample droplets may have only a single sample molecule. This improves the ionization process.

The specimen may comprise, for example, polar lipids, and the vapor or aerosol may comprise aqueous droplets converted to polar lipids.

The specimen may comprise triglyceride.

The sample supplied with the matrix may include ionized sample molecules.

The method may further include the step of generating a gas phase sample, a vapor sample, an aerosol, or a liquid from the sample being analyzed.

The vapor phase sample, vapor sample, or aerosol may be generated by heating the sample containing the sample, or by diathermy evaporation of the sample.

The method may be either a surgical method or part of a non-surgical method. For example, the method may be a surgical method in which the sample may be human or animal tissue containing the sample. The sample may undergo electrosurgical diathermy evaporation, or other form of rapid evaporation, to form a gas phase sample, vapor sample, or aerosol. As an example, devices and methods may be used to identify human cells in breast cancer surgery. By analyzing the sample ions, it is possible to determine whether or not the tissue is cancerous.

Alternatively, the method may include a non-surgical method. For example, human or animal tissue that is not part of the human or animal body (ie, pre-extracted, placed, or removed) may be analyzed, or a sample or biological tissue other than human or animal tissue is analyzed. You may. Also, by analyzing the sample ions, it is possible to determine the characteristics or components of the sample, such as whether or not the sample contains cancer tissue.

Embodiments include country of origin identification, drug testing, food safety testing (eg dairy products), cosmetic testing, military use, air pollution testing, post-mortem analysis, microbial identification (eg bacteria), and other non-surgical embodiments such as automated sampling. It may be used in the method.

The method may be used to analyze abiotic samples and compounds.

The sample formed from the sample may be partially charged and / or may have a relatively high organic content.

The device may further include evaporating the matrix from the sample in the second smaller dissolved sample droplet so as to provide sample ions substantially separated from the matrix.

The step of evaporating the matrix from the sample may cause the separation of the dissolved sample ions ionically bonded from the matrix to form gas phase sample ions.

After the step of evaporating the matrix from the sample, the method may further include capturing the sample ions in an ion trap and / or guiding the sample ions using an ion guide.

The method may further include analyzing the sample ions.

The steps to analyze the sample ions are (i) mass spectrometry of the sample ions, (ii) analyzing the ion mobility or different ion mobility of the sample ions, and (iii) the ion cross section or collision cross section of the sample ions. Analyzing, (iv) separating the sample ions according to their ion mobility or different ion mobility, (v) before mass spectrometrically analyzing the sample ions, the sample ions have their ion mobility or different ion mobility. Separation according to, or (vi) exclusion or disposal of sample ions based on their ion conductivity or different ion conductivity may further be included.

The matrix may be fed to the specimen, which is in the gas phase, in the form of vapor, in the form of an aerosol, or in the liquid phase.

The step of feeding the matrix compound to the sample may include feeding the matrix molecules to the sample and intermixing the matrix molecules with the sample, but the matrix is in the gas phase.

The mixture of sample and matrix is from the high pressure region to the low pressure region so that the gas phase matrix cools and condenses into the liquid and the sample dissolves in the liquid matrix to form the first dissolved sample droplets. It may be transferred.

The matrix comprises (i) a solvent for specimens, smoke, sprays, liquids, gases, surgical smoke, aerosols, or vapors, (ii) organic solvents, (iii) volatile compounds, (iv) polar molecules, (v). It may be selected from the group consisting of water, (vi) one or more alcohols, (vii) methanol, (viii) ethanol, (ix) isoprapanol, (x) acetone, and (xi) acetonitrile.

The step of colliding the first dissolved sample droplet with the collision surface or gas may further include accelerating the first dissolved sample droplet into the collision surface or gas.

The step of accelerating the first dissolved sample droplet to the collision surface or gas also includes using a pressure difference to accelerate the first dissolved sample droplet into the collision surface or gas. Good.

The method may further include analyzing the sample ions using a mass spectrometer equipped with an atmospheric interface adjacent to the vacuum chamber, where the first dissolved sample droplets collide with the pressure difference across the atmospheric interface. Or it is accelerated into the gas.

The method may further include heating the collision surface (or otherwise the collision gas).

The methods are (i) about <100 ° C., (ii) about 100-200 ° C., (iii) about 200-300 ° C., (iv) about 300-400 ° C., (v) about 400-500 ° C., (vi). About 500-600 ° C, (vii) about 600-700 ° C, (viii) about 700-800 ° C, (ix) about 800-900 ° C, (x) about 900-1000 ° C, (xi) about 1000-1100 ° C , And (xii) may further include heating the collision surface (or otherwise the collision gas) to a temperature selected from the group consisting of about> 1100 ° C.

The matrix may be supplied to the specimen by a matrix conduit.

The method may further include analyzing sample ions using an ion analyzer located downstream of the outlet of the matrix conduit.

The distance x between the outlet of the matrix conduit and the inlet of the ion analyzer is (i) about 0.1 to 0.5 mm, (ii) about 0.5 to 1.0 mm, and (iii) about 1. 0 to 1.5 mm, (iv) about 1.5 to 2.0 mm, (v) about 2.0 to 2.5 mm, (vi) about 2.5 to 3.0 mm, (vi) about 3.0 to 3.5 mm, (viii) about 3.5 to 4.0 mm, (ix) about 4.0 to 4.5 mm, (x) about 4.5 to 5.0 mm, (xi) about 5.0 to 5. 5 mm, (xxii) about 5.5-6.0 mm, (xxii) about 6.0-6.5 mm, (xiv) about 6.5-7.0 mm, (xv) about 7.0-7.5 mm, (Xvi) about 7.5-8.0 mm, (xvii) about 8.0-8.5 mm, (xvii) about 8.5-9.0 mm, (xx) about 9.0-9.5 mm, (xx) ) About 9.5 to 10.0 mm, (xxii) about 0.1 to 10 mm, (xxii) about 0.1 to 7.5 mm, (xxiii) about 0.1 to 5.1 mm, (xxiv) about 0. It may be selected from the group consisting of 5 to 5.1 mm and (xxv) about 0.5 to 5.0 mm.

The apparatus includes (i) about 50 to 100 μl / min, (ii) about 100 to 150 μl / min, (iii) about 150 to 200 μl / min, (iv) about 200 to 250 μl / min, and (v) about 250 to 300 μl. / Min, (vi) about 300-350 μl / min, (vii) about 350-400 μl / min, (viii) about 400-450 μl / min, (ix) about 450-500 μl / min, (x) about 500-550 μl / Min, (xi) about 550-600 μl / min, (xii) about 600-650 μl / min, (xiii) about 650-700 μl / min, (xiv) 700-750 μl / min, (xv) about 750-800 μl / min, (xvi) about 800-850 μl / min, (xvii) about 850-900 μl / min, (xvii) about 900-950 μl / min, (xix) about 950-1000 μl / min, (xx) about 50 μl / min ~ Specimens are fed by a matrix conduit at a flow rate selected from the group consisting of 1 ml / min, (xxxi) about 100-800 μl / min, (xxxi) about 150-600 μl / min, and (xxii) about 200-400 μl / min. You may.

The outlet of the matrix conduit may be on the opposite side of the inlet of the ion analyzer or may be coaxial with the inlet of the ion analyzer.

The method may further include mass and / or ion mobility analysis of sample ions with an ion sampler to obtain sample ion data, the method analyzing rock mass, lock mobility, or calibration ions. It may further include calibrating the ion analyzer based on the data obtained from analyzing the rock mass, rock mobility, or calibration ions, or adjusting the sample ion data.

According to another aspect, methods of mass and / or ion mobility analysis are provided, including methods as described above.

According to another aspect
The sample inlet for receiving the sample and
A mixing region for mixing the sample with the matrix compound so that the sample dissolves in the matrix during use to form a first dissolved sample droplet.
With a collision surface or gas,
The first dissolved sample droplet is arranged and adapted to collide with a collision surface or gas so that the first dissolved sample droplet breaks into a plurality of second smaller dissolved sample droplets. A device for performing rapid evaporation ionized mass spectrometry (“REIMS”) is provided with the device.

The device may further comprise a device adapted to evaporate the matrix from a second smaller dissolved sample droplet so as to provide the sample ions separated from the matrix.

The device further comprises a device adapted to evaporate the matrix from the sample such that the charge is transferred to or from the sample to ionize the sample to form sample ions. May be good.

The device may further include an ion trap and / or an ion guide.

In order for the device to capture the sample ions in an ion trap and / or guide the sample ions using an ion guide, the matrix is further equipped with a adapted device that is placed after evaporating from the sample during use. May be good.

The device may further include an analyzer for analyzing sample ions.

The analyzers are (i) mass spectrometers for mass spectrometry of sample ions, (ii) ion mobility or different ion mobility analyzers, and (iii) analysis for analyzing ion cross sections or collision cross sections of sample ions. Vessel, (iv) Analyzer for separating sample ions according to their ion mobility or different ion mobility, (v) Specimens according to their ion mobility or different ion mobility before mass spectrometry. Further may be provided with a separator for separating ions, or a device adapted to eliminate or discard (vi) sample ions based on their ion conductivity or different ion conductivity.

The matrix may be fed to the specimen at the time of use, but the specimen is in the gas phase, in the form of vapor, in the form of an aerosol, or in the liquid phase.

The device may further comprise a device adapted to feed the matrix molecules to the sample and intermix the matrix molecules with the sample, but the matrix is in the gas phase.

The device cools the gas phase matrix, condenses it into a liquid, and dissolves the sample-matrix mixture in the liquid matrix to form a first dissolved sample droplet from the high pressure region to the low pressure region. Further devices may be provided that are arranged and adapted for transfer to the area.

The matrix comprises (i) a solvent for specimens, smoke, sprays, liquids, gases, surgical smoke, aerosols, or vapors, (ii) organic solvents, (iii) volatile compounds, (iv) polar molecules, (v). It may be selected from the group consisting of water, (vi) one or more alcohols, (vii) methanol, (viii) ethanol, (ix) isoprapanol, (x) acetone, and (xi) acetonitrile.

According to another aspect, a mass and / or ion mobility analyzer is provided with the equipment as described above.

The analyzer may further include a device adapted and arranged to accelerate the first dissolved sample droplet into the collision surface or gas.

The analyzer may further include a device adapted and arranged to hold a pressure difference to accelerate the first dissolved sample droplet into the collision surface or gas.

The analyzer may further include an analyzer arranged to analyze the sample ions, the analyzer further comprises an atmospheric interface adjacent to the vacuum chamber, and the first dissolved sample droplets are atmospheric. The pressure difference across the interface accelerates into the collision surface or gas.

The analyzer may further include a heater for heating the collision surface (or otherwise the collision gas).

The heaters are (i) about <100 ° C, (ii) about 100-200 ° C, (iii) about 200-300 ° C, (iv) about 300-400 ° C, (v) about 400-500 ° C, (vi). ) About 500 to 600 ° C, (vii) about 600 to 700 ° C, (viii) about 700 to 800 ° C, (ix) about 800 to 900 ° C, (x) about 900 to 1000 ° C, (xi) about 1000 to 1100 It may be arranged to heat the collision surface (or otherwise the collision gas) to a temperature selected from the group consisting of ° C. and (xi) about> 1100 ° C.

The analyzer may further include a matrix conduit for supplying the matrix to the specimen.

The analyzer may further include an ion analyzer for analyzing sample ions, which is located downstream of the outlet of the matrix conduit.

The distance x between the outlet of the matrix conduit and the inlet of the ion analyzer is (i) about 0.1 to 0.5 mm, (ii) about 0.5 to 1.0 mm, and (iii) about 1. 0 to 1.5 mm, (iv) about 1.5 to 2.0 mm, (v) about 2.0 to 2.5 mm, (vi) about 2.5 to 3.0 mm, (vi) about 3.0 to 3.5 mm, (viii) about 3.5 to 4.0 mm, (ix) about 4.0 to 4.5 mm, (x) about 4.5 to 5.0 mm, (xi) about 5.0 to 5. 5 mm, (xxii) about 5.5-6.0 mm, (xxii) about 6.0-6.5 mm, (xiv) about 6.5-7.0 mm, (xv) about 7.0-7.5 mm, (Xvi) about 7.5-8.0 mm, (xvii) about 8.0-8.5 mm, (xvii) about 8.5-9.0 mm, (xx) about 9.0-9.5 mm, (xx) ) About 9.5 to 10.0 mm, (xxii) about 0.1 to 10 mm, (xxii) about 0.1 to 7.5 mm, (xxiii) about 0.1 to 5.1 mm, (xxiv) about 0. It may be selected from the group consisting of 5 to 5.1 mm and (xxv) about 0.5 to 5.0 mm.

The analyzers are (i) about 50-100 μl / min, (ii) about 100-150 μl / min, (iii) about 150-200 μl / min, (iv) about 200-250 μl / min, (v) about 250-. 300 μl / min, (vi) about 300-350 μl / min, (vii) about 350-400 μl / min, (viii) about 400-450 μl / min, (ix) about 450-500 μl / min, (x) about 500- 550 μl / min, (xi) about 550-600 μl / min, (xii) about 600-650 μl / min, (xiii) about 650-700 μl / min, (xiv) 700-750 μl / min, (xv) about 750-800 μl / Min, (xvi) about 800-850 μl / min, (xvii) about 850-900 μl / min, (xvii) about 900-950 μl / min, (xix) about 950-1000 μl / min, (xx) about 50 μl / min Matrix via matrix conduit at a flow rate selected from the group consisting of ~ 1 ml / min, (xxii) about 100-800 μl / min, (xxii) about 150-600 μl / min, and (xxii) about 200-400 μl / min. May be further equipped with a pump for supplying the sample.

The outlet of the matrix conduit may be on the opposite side of the inlet of the ion analyzer or may be coaxial with the inlet of the ion analyzer.

The mass spectrometer may further include a mass spectrometer for analyzing sample ions to obtain sample ion data, which analyzes and locks rock mass, lock mobility, or calibration ions. The ion analyzer may be calibrated based on the data obtained from analyzing the mass, lock mobility, or calibrated ions, or may be further arranged to adjust the sample ion data.

According to another aspect
Contacting biological tissue with electrosurgical equipment and activating electrosurgical equipment to generate specimens, smoke, sprays, liquids, gases, surgical smoke, aerosols, or vapors.
Inhaling specimens, smoke, sprays, liquids, gases, surgical smoke, aerosols, or vapors,
Mixing the matrix with aspirated specimens, smoke, sprays, liquids, gases, surgical smoke, aerosols, or vapors,
Aspirated specimens, smoke, sprays, liquids, gases, surgical smokes, aerosols, or vapors bound to the matrix may be placed in the vacuum chamber of the mass spectrometer to form specimen ions (or collision surfaces). As an alternative, it is possible to collide with a collision gas)
Methods of electrosurgery are provided that include mass and / or ion mobility analysis of sample ions.

According to another aspect
Rapid Evaporation Ionized Mass Spectrometry (“REIMS”) electrosurgical instruments with one or more electrodes,
Electrosurgical tools have been arranged and adapted to activate electrosurgical tools upon contact with biological tissue during use, such as to generate specimens, smoke, sprays, liquids, gases, surgical smoke, aerosols, or vapors. With the device
With devices adapted to inhale specimens, smoke, sprays, liquids, gases, surgical smoke, aerosols, or vapors,
With a device adapted to mix the matrix with the sample, smoke, spray, liquid, gas, surgical smoke, aerosol, or vapor.
(I) Specimens, smoke, sprays, liquids, gases, surgical smoke, aerosols, or vapors are arranged to collide with the collision surface (or otherwise the collision gas) so as to form specimen ions during use. Mass and / or ion mobility analysis provided with a collision surface (or otherwise collision gas) that may be placed in the vacuum chamber of the mass spectrometer, and (ii) a mass spectrometer for mass spectrometry of sample ions. An electrosurgical device with a meter is provided.

According to one embodiment, the matrix may first be fed as a solid, eg, a powder, sublimated or melted and evaporated to form the matrix in a vapor or vapor phase intermixed with the sample.

Alternatively, the matrix may be fed to the specimen as a liquid, aerosol, or vapor, or may be intermixed with the specimen. If the specimen and / or matrix is in the form of a liquid, the mixture of specimen and matrix may need to be subsequently converted into a first dissolved specimen droplet, for example by jetting.

The permittivity of the matrix can be very high such that the solvated ions of the sample are present in the condensed phase due to the solvation of the sample involved in ion dissolution. In these cases, a collision with the collision surface (or otherwise the collision gas) is likely to produce solvate ions in the gas phase, which ultimately (negative ion mode, ie [MH]. ^It may give rise to ions formed by deprotonation (at ⁻ ), ions formed by protonation (in positive ion mode, ie [M + H] ⁺ ), and / or molecular ions.

Isoprapanol is a particularly convenient matrix used, for example, for lipid species.

As an example, for a sample with polar lipids, the matrix may be a low molecular weight alcohol (eg methanol, ethanol, isopropanol) or a ketone (eg acetone), or a low molecular weight alcohol (eg methanol, ethanol, isopropanol). Alternatively, a ketone (for example, acetone) may be provided. These matrices have been shown to enhance the ionization of all or certain species that would otherwise be detected as low intensity and no vapor in the matrix.

The mixture of sample and matrix may be a homogeneous mixture or a heterogeneous mixture.

A voltage may be applied to the ion trap or ion guide to capture or guide the ions, respectively. The ions may then be delivered to the ion analyzer from an ion trap or ion guide to analyze the mass and / or ion mobility of the ions.

Ions may be separated according to ion mobility prior to mass spectrometry. Ions may then be eliminated or discarded based on their ion mobility.

Any range of the above ranges may be combined with any range in the list of ranges for distance x.

The inlet of the ion analyzer may be an opening or an orifice that separates the vacuum chamber of the ion analyzer from the higher pressure region upstream of the ion analyzer. For example, the inlet may be an atmospheric pressure interface.

According to an alternative embodiment, the matrix conduit may deliver the matrix directly into the sample transfer tube performing the step of providing the sample.

Alternatively, a sample transfer tube may be provided to perform the step of providing the sample, and the outlet of the matrix conduit may be provided at a location around the sample transfer tube. The gas stream may be arranged to clear the matrix from the outlet to the inlet of the ion analyzer that analyzes the ions.

The device for evaporating the sample may be equipped with an electrosurgical device such as a diathermy device.

The device may have an end, point or region for insertion onto the sample to evaporate the sample, with the sample inlet adjacent to the end, point or region.

The device may include a source of matrix compounds for supplying the matrix compounds to the conduit.

The accelerating means is between the two regions in order to accelerate the first dissolved sample droplet between the first region and the second region to the collision surface (or otherwise the collision gas). A vacuum pump may be provided to generate the pressure difference.

The device may include a mass spectrometer with an atmospheric interface located between the first and second regions, the second region being connected to a vacuum pump and the collision surface (or alternative). A vacuum chamber for accommodating the collision gas) may be provided.

The device may include an ion trap or ion guide to capture or guide the sample ions.

The ion analyzer may include a mass and / or ion mobility analyzer or analyzer.

The device may be arranged and adapted to perform any of the methods described herein.

The mixing region may be provided upstream of the inlet of the ion analyzer, or the mixing region may be provided at least a portion downstream of the ion analyzer.

The inlet of the ion analyzer may be an opening or an orifice that separates the vacuum chamber of the ion analyzer from the higher pressure region upstream of the ion analyzer. For example, the inlet may be equipped with an atmospheric pressure interface.

The matrix conduit may deliver the matrix directly to the sample transfer tube, performing the steps of providing the sample.

Alternatively, a sample transfer tube may be provided to perform the step of providing the sample, and the outlet of the matrix conduit may be provided at a location around the sample transfer tube. The gas stream may be arranged to clear the matrix from the outlet to the inlet of the ion analyzer that analyzes the ions.

It may include rock mass, lock mobility, or calibration compound ions or sources of ions.

The rock mass, lock mobility or calibration compound / ion may be introduced into the matrix conduit, sample conduit, or fed into a separation tube.

According to the embodiment, the aerosol particles containing the sample (or gas phase sample molecule) may be introduced into the mass spectrometer together with the volatile matrix compound, and the volatile matrix compound is an organic solvent. It is also good. The volatile matrix compound may be introduced into the specimen as a solid (eg, powder), liquid, aerosol, or vapor. The mixture of sample and matrix may be aspirated into the mass spectrometer by the pressure difference from the inlet to the analyzer. The low pressure inside the mass spectrometer provides a gas that mixes the expanding sample and matrix, causing the temperature in the free jet region to drop. This causes the gas or evaporated specimen and / or matrix to condense, causing the specimen to dissolve in the matrix. The role of the matrix compound may be to generate aerosol particles that contain a matrix that exceeds the sample molecule and incorporate the sample molecule in the form of a solvent. Solventalization substantially removes the intermolecular secondary binding force between the sample molecules as each dissolved sample molecule is completely surrounded by the matrix molecules. Separation of the sample molecules within the condensed phase increases the probability that the aerosol particles will form clusters, each containing only a single sample molecule, when the aerosol particles collide with the collision surface. Matrix molecules may have a high dielectric constant and / or a high vapor pressure.

Next, as an example only, various embodiments will be described with reference to the accompanying drawings.

Rapid evaporation ionized mass spectrometry where a DC voltage is applied to the bipolar forceps that result in the production of aerosols or surgical fumes, which are captured through the cleaning section of the bipolar forceps and then transferred to a mass spectrometer for ionization and mass spectrometry. It is a figure which shows the method of ("REIMS"). It is a figure which shows one mounting form in which a sample and a matrix may be provided in a gas phase or a vapor phase. FIG. 5 illustrates another embodiment in which a sample and a matrix may be provided in the liquid phase. It is a figure which shows the mass spectrum obtained without using a matrix. It is a figure which shows the mass spectrum obtained by using a matrix. It is a figure which shows one Embodiment of the mass spectrometer interface which comprises a Venturi device for introducing a sample aerosol and a matrix into a mass spectrometer. It is an enlarged view of FIG. 5B. It is a proximity view of the sampling device in the interface. FIG. 5 shows how the ion signal detected using the embodiment of FIG. 5 changes depending on the distance between the outlet of the matrix conduit and the inlet of the ion analyzer. It is a figure which shows how the ion signal detected by using the embodiment of FIG. 5 changes depending on the flow rate of a matrix. It is a figure which shows the mass spectrum obtained by using the matrix flow rate of different isoprapanols. It is a figure which shows the mass spectrum obtained by using the matrix flow rate of different isoprapanols. It is a figure which shows the mass spectrum obtained by using the matrix flow rate of different isoprapanols. It is a figure which shows the mass spectrum obtained by using the matrix flow rate of different isoprapanols. It is a figure which shows the mass spectrum obtained by using the matrix flow rate of different isoprapanols. It is a figure which shows the mass spectrum obtained by using the matrix flow rate of different isoprapanols. It is a figure which shows the mass spectrum obtained by using the matrix flow rate of different isoprapanols. It is a figure which shows the mass spectrum obtained by using the matrix flow rate of different isoprapanols. It is a figure which shows the mass spectrum obtained by using the matrix flow rate of different isoprapanols. It is a figure which shows the mass spectrum obtained by using a rock mass compound. It is a figure which shows the mass spectrum acquired without using a rock mass compound. It is a figure which shows the result of the principal component analysis on the data acquired over different days with and without the use of rock mass ion. It is a figure which shows the result of the principal component analysis on the data acquired for different tissue types with and without the use of rock mass ion. It is a figure which shows one Embodiment which the collision surface is a sphere. It is a figure which shows one Embodiment which the collision surface has a coil shape. It is a figure which shows the mass spectrum obtained by using the collision surface which is not heated. It is a figure which shows the mass spectrum obtained by using the heated collision surface. It is a figure which shows the mass spectrum acquired when the sample which IPA matrix was introduced upstream of the heated collision surface was analyzed. FIG. 5 shows mass spectra obtained from the same analysis, except when the matrix is not used. It is a figure which showed the mass spectrum when the collision surface was not overheated and the matrix was not used, which was obtained from the same analysis. FIG. 5 shows the total ion currents detected for several different distances between the outlet of a matrix conduit having an inner diameter of 250 μm and the inlet of a mass spectrometer into a vacuum chamber. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 5 shows the total ion currents detected for several different distances between the outlet of a matrix conduit having an inner diameter of 100 μm and the inlet of a mass spectrometer into a vacuum chamber. FIG. 16A shows mass spectra obtained at different distances in FIG. 16A. FIG. 16A shows mass spectra obtained at different distances in FIG. 16A. FIG. 16A shows mass spectra obtained at different distances in FIG. 16A. FIG. 16A shows mass spectra obtained at different distances in FIG. 16A. FIG. 16A shows mass spectra obtained at different distances in FIG. 16A. FIG. 16A shows mass spectra obtained at different distances in FIG. 16A. FIG. 16A shows mass spectra obtained at different distances in FIG. 16A. FIG. 16A shows mass spectra obtained at different distances in FIG. 16A. FIG. 5 shows the total ion currents detected for several different distances between the outlet of a matrix conduit having an inner diameter of 50 μm and the inlet of a mass spectrometer into a vacuum chamber. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. FIG. 15A shows mass spectra acquired at different distances in FIG. 15A. It is a figure which shows the spectrum acquired for the matrix conduit which has an inner diameter of 50 μm. It is a figure which shows the spectrum acquired for the matrix conduit which has an inner diameter of 100 μm. It is a figure which shows the spectrum acquired for the matrix conduit which has an inner diameter of 250 μm. FIG. 5 shows the total ion currents detected for several different distances between the outlet of a matrix conduit having an inner diameter of 250 μm and the inlet of a mass spectrometer into a vacuum chamber. FIG. 19A shows mass spectra obtained at different distances. FIG. 19A shows mass spectra obtained at different distances. FIG. 19A shows mass spectra obtained at different distances. FIG. 19A shows mass spectra obtained at different distances. FIG. 19A shows mass spectra obtained at different distances. FIG. 19A shows mass spectra obtained at different distances. FIG. 19A shows mass spectra obtained at different distances. FIG. 5 shows the total ion currents detected for several different distances between the outlet of a matrix conduit having an inner diameter of 100 μm and the inlet of a mass spectrometer into a vacuum chamber. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 20A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 20A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 20A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 20A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 20A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 20A. FIG. 5 shows the total ion currents detected for several different distances between the outlet of a matrix conduit having an inner diameter of 50 μm and the inlet of a mass spectrometer into a vacuum chamber. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 21A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 21A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 21A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 21A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 21A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 21A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 21A. It is a figure which shows the mass spectrum acquired at the different distance of FIG. 21A. It is a figure which shows the mass spectrum acquired in the negative ion mode without introducing a matrix into a sample flow, and without using a collision surface. It is a figure which shows the mass spectrum acquired without using the collision surface by introducing the matrix into the sample flow. It is a figure which shows the mass spectrum obtained by introducing a matrix into a sample flow, and using a collision plane. It is a figure which shows the mass spectrum acquired in the negative ion mode without introducing a matrix into a sample flow, and without using a collision surface. It is a figure which shows the mass spectrum acquired without using the collision surface by introducing the matrix into the sample flow. It is a figure which shows the mass spectrum obtained by introducing a matrix into a sample flow, and using a collision plane. It is a figure which shows the mass spectrum obtained from the analysis of the normal breast tissue without using a matrix. It is a figure which shows the mass spectrum obtained from the analysis of the normal breast tissue using a matrix. It is a figure which shows the analysis method including constructing the classification model by various embodiments. It is a figure which shows the set of standard sample mass spectra obtained from two classes of known standard samples. It is a diagram showing a multivariate space having three dimensions defined by an intensity axis, the multivariate space has a plurality of reference points, and each reference point is a set of three peak intensity values drawn from a standard sample mass spectrum. Correspond. It is a figure which shows the general relationship between the cumulative variance and the number of components of a PCA model. FIG. 6 shows a PCA space having two dimensions defined by a principal component axis, the PCA space containing a plurality of transformed reference points or scores, each transformed reference point corresponding to the reference point of FIG. .. It is a diagram showing a PCA-LDA space having one dimension or an axis, the LDA is executed based on the PCA space of FIG. 29, and the PCA-LDA space contains a plurality of further converted reference points or class scores, respectively. Further converted reference points correspond to the converted reference points or scores in FIG. It is a figure which shows the analysis method including using the classification model by various embodiments. It is a figure which shows the sample mass spectrum obtained from an unknown sample. It is a figure which shows the PCA-LDA space of FIG. 30, and the PCA-LDA space further includes the projected sample point of PCA-LDA extracted from the peak intensity value of the sample mass spectrum of FIG. It is a figure which shows the analysis method including constructing the classification library by various embodiments. It is a figure which shows the analysis method including using the classification library by various embodiments.

More details below describe various embodiments that generally relate to the generation of aerosols, surgical smoke, or vapors from one or more regions of a target (eg, tissue in the body) using an ambient ionized ion source. explain.

The aerosol, surgical smoke, or vapor is then mixed with the matrix and sucked into the vacuum chamber of the mass spectrometer. When the mixture collides with the collision surface, the aerosol, smoke, or vapor is ionized by the collision ionization, causing the generation of sample ions.

The resulting sample ions (or debris or generated ions extracted from the sample ions) may then be analyzed for mass and / or ion mobility, and the resulting mass and / or ion mobility analysis data will then be analyzed in real time. Multivariate analysis may be performed to determine the properties of one or more of the targets.

For example, a multivariate analysis may be able to determine if some of the tissue currently being resected is cancerous.

Peripheral Ionized Ion Sources According to various embodiments, the device is used to generate aerosols, smoke, or vapors from one or more areas of a target (eg, tissue in the body). The device may include, for example, an ambient ionized ion source characterized by the ability to generate a sample aerosol, smoke, or vapor from a natural or unmodified target. Aerosols, smoke, or vapors may then be mixed with the matrix and aspirated into the vacuum chamber of the mass and / or ion mobility analyzer. The mixture is struck by the collision surface and the aerosol, smoke, or vapor is ionized by slamming ionization, causing the generation of sample ions. The resulting sample ions (or debris or generated ions extracted from the sample ions) may then be analyzed for mass and / or ion mobility, and the resulting mass and / or ion mobility analysis data will be targeted in real time. Multivariate analysis or other mathematical processing may be performed to determine one or more properties of. For example, a multivariate analysis may be able to determine if some of the tissue currently being resected is cancerous.

The need to add the matrix or reagents directly to the sample makes it impossible to perform tissue analysis in the body and, more generally, to provide a quick and simple analysis of the target material. Will be understood.

Thus, conversely, the peri-ionization technique is particularly favorable, firstly, the peri-ionization technique does not require the addition of a matrix or reagent to the sample (and is therefore suitable for analysis of tissues in the body), and secondly. Peripheral ionization techniques can perform rapid and simple analysis of the target material. Other types of ionized ion sources, such as matrix-assisted laser desorption / ionization (“MALDI”) ion sources, require the matrix or reagent to be added to the sample prior to ionization.

A number of different ambient ionization techniques are known and are intended to fall within the scope of the present invention. As a historical record, desorption electrospray ionization (“DESI”) was the first ambient ionization technique developed and was disclosed in 2004. Since 2004, a number of other ambient ionization techniques have been developed. These ambient ionization techniques differ in their rigorous ionization methods, but they have the same general function as the ability to generate gas phase ions directly from natural (ie, untreated or unmodified) samples. Share. A special advantage of the various ambient ionization techniques intended to fall within the scope of the present invention is that the various ambient ionization techniques do not require any prior sample formulation. As a result, various ambient ionization techniques can analyze both in-vivo and extracorporeal tissue samples without the time and expense of adding matrices or reagents to the tissue samples or other target materials. it can.

A list of ambient ionization techniques intended to fall within the scope of the present invention is provided in the table below.

According to one embodiment, the ambient ionization ion source may include a rapid evaporative ionization mass spectrometry (“REIMS”) ion source, one or more for producing an aerosol, or Joule heat, to generate a blast of surgical smoke. A DC voltage is applied to the electrodes of.

However, it will be understood that other ambient ion sources, including those mentioned above, may also be utilized. For example, according to another embodiment, the ambient ionization ion source may include a laser ablation ion source. According to one embodiment, the laser ionization ion source may include a mid-infrared laser ionization ion source. For example, there are several lasers that emit radiation at or near 2.94 μm, which corresponds to a peak in the water absorption spectrum. According to various embodiments, the ambient ionization ion source may include a laser ablation ion source having a wavelength of 2.94 μm and close to 2.94 μm based on the high water absorption coefficient of water. According to one embodiment, the laser ablation ion source may include an Er: YAG laser that emits radiation at 2.94 μm.

In another embodiment, a mid-infrared optical parametric oscillator (“OPO”) may be used to generate a laser ablation ion source with wavelengths greater than 2.94 μm. For example, a ZGP-OPO pumped with Er: YAG may be used to generate a laser irradiation line having a wavelength of, for example, 6.1 μm, 6.45 μm, or 6.73 μm. In some situations, it may be advantageous to use a laser ablation ion source with wavelengths shorter or longer than 2.94 μm, as only the surface layer is cut and less likely to cause thermal damage. According to one embodiment, a Co: MgF2 laser may be used as the laser ablation ion source and the laser may be adjusted to 1.75 to 2.5 μm. According to another embodiment, an optical parametric oscillator (“OPO”) system pumped by an Nd: YAG laser may be used to generate a laser ablation ion source with a wavelength of 2.9-3.1 μm. Good. According to another embodiment, a CO2 laser having a wavelength of 10.6 μm may be used to generate an aerosol, smoke, or vapor.

According to other embodiments, the ambient ionization ion source may comprise an ultrasonic ablation ion source, or hybrid electrosurgical, i.e., an ultrasonic ablation source, where the ultrasonic ablation source produces a liquid sample and the liquid sample It is then aspirated as an aerosol. The ultrasonic ablation ion source may include concentrated or defocused ultrasonic waves.

According to one embodiment, the first device for generating aerosols, smoke, or vapors from one or more regions of the target may include tools that utilize DC voltage, such as continuous RF waveforms. .. According to other embodiments, radio frequency tissue anatomical systems arranged to deliver pulsed plasma RF energy to the instrument may be used. The tool may include, for example, PlasmaBlade®. Pulse plasma RF instruments operate at lower temperatures than conventional electrosurgical instruments (eg, 200-350 ° C compared to 40-170 ° C), thereby reducing the depth of burns. The pulse waveform and duty cycle may be used for both cutting and working solidification modes by inducing an electrical plasma along the cutting edge (s) of the thin insulator electrode.

According to one embodiment, the first device is an excision device, such a device synthesized with any other device herein, an electrosurgical argon plasma coagulation device, a hybrid argon plasma coagulation and a water / saline jet. It is equipped with a surgical water / saline jet device such as a device.

In another embodiment, it is contemplated that the first device for generating aerosols, smoke, or vapors from the target may include an argon plasma coagulation ("APC") device. Argon plasma coagulation devices use a jet of ionized argon gas (plasma) directed through a probe. The probe may pass through the endoscope. Argon plasma coagulation is basically a non-contact process, like a probe placed a short distance from the target. Argon gas is released from the probe and then ionized by a high pressure discharge (eg 6 kV). A high frequency current is then transmitted through the jet of gas, resulting in solidification of the target at the other end of the jet. The depth of solidification is usually only a few millimeters.

The first device, surgical or electrosurgical device, the device or probe or other sampling device or probe disclosed in any aspect or embodiment herein is one or more hydraulic surgical devices, surgical water jet devices, Non-contact surgical devices such as argon plasma coagulation devices, hybrid argon plasma coagulation devices, water jet devices, and laser devices may be provided.

Non-contact surgical devices are defined as surgical devices that are arranged and adapted to cut, crush, liquefy, aspirate, discharge, or otherwise destroy biological tissue without physical contact with the tissue. May be done. Examples include laser devices, hydraulic surgery devices, argon plasma coagulation devices, and hybrid argon plasma coagulation devices. Treatment may appear to be relatively safe, as non-contact devices do not have to make physical contact with tissue, to treat delicate tissue with low adhesion within the tissue, such as skin or fat. Can be used.

Rapid evaporation ionization mass spectrometry ("REIMS")
FIG. 1 shows a method of rapid evaporation ionization mass spectrometry (“REIMS”), where bipolar forceps 1 may be brought into contact with tissue 2 in the body of patient 3. In the example shown in FIG. 1, bipolar forceps 1 may be brought into contact with patient 3's brain tissue 2 during the course of surgery in the patient's brain. A DC voltage may be applied to the bipolar forceps 1 from the DC voltage generator 4, and the bipolar forceps 1 localize the Joule heat or diathermy heat of the tissue 2. As a result, aerosol or surgical smoke 5 is generated. Aerosol or surgical smoke 5 may then be captured or otherwise aspirated through the wash port of bipolar forceps 1. Therefore, the cleaning port of the bipolar forceps 1 is reused as a suction port. The aerosol or surgical smoke 5 may then be passed from the wash (suction) port of the bipolar forceps 1 into a tube 6 (eg, a 1/8 inch or 3.2 mm diameter Teflon tube). The tube 6 is arranged to transfer the aerosol or surgical smoke 5 to the atmospheric pressure interface 7 of the mass spectrometer 8.

According to various embodiments, a matrix comprising an organic solvent such as isopranol may be added to the aerosol or surgical smoke 5 at atmospheric pressure interface 7. The mixture of the aerosol 3 and the organic solvent may then be arranged in the vacuum chamber of the mass spectrometer 8 so as to collide with the collision surface. According to one embodiment, the collision surface may be heated. Aerosols are ionized when they collide with the collision surface, resulting in the generation of sample ions. The ionization efficiency of generating sample ions may be improved by the addition of an organic solvent (ie, matrix).

Specimen ions generated by hitting the aerosol, smoke, or steam 5 against the collision surface are then passed to the next stage of the mass and / or ion mobility analyzer and mass within the mass and / or ion mobility analyzer. And / or undergo ion mobility analysis. The mass spectrometer may include, for example, a quadrupole mass spectrometer or a time-of-flight mass spectrometer.

FIG. 2 shows a schematic view of one embodiment. The device may include an ion analyzer 207 having ion optics 212 such as an inlet 206, a vacuum region 208, a collision surface 209, and a Stepwave® ion guide located within the vacuum region 208. The device may also include a sample transfer tube 202 and a matrix conduit 203. The sample transfer tube 202 is the inlet for receiving the aerosol sample 201, which may correspond to the aerosol or surgical smoke 5 described with respect to FIG. 1, from the sample being investigated, and the flow of the ion analyzer 207. It has an outlet connected to inlet 206. The matrix conduit 203 has an inlet for receiving the matrix compound and an outlet that intersects the sample transfer tube 202 so that the matrix 204 can intermix with the aerosol sample 201 within the sample transfer tube 202. The T-shaped joint component may be provided at the point of contact between the tubes 202, 203 and 206. The tubes 202, 203, and 206 may be removably inserted into the T-joint.

Next, the operation method of the device of FIG. 2 will be described. Samples, such as biological samples, may undergo REIMS techniques. For example, a diathermy device may be used to evaporate the biological tissue from the sample to form an aerosol, eg, as described above with respect to FIG. The aerosol particles 201 are then introduced into the inlet of the sample transfer tube 202. Matrix compound 204 is introduced into the inlet of matrix conduit 203. The aerosol particles 201 and the matrix compound 204 are attracted toward the inlet 206 of the ion analyzer 207 by the pressure difference created by the vacuum chamber 208, which is lower pressure than the inlets to the tubes 202, 203. Aerosol particles 201 may encounter molecules of matrix compound 204 in the region where the sample transfer tube 202 intersects the matrix conduit 203 and downstream of the region where the sample transfer tube 202 intersects the matrix conduit 203. The aerosol particles 201 are intermixed with the matrix 204 to form aerosol particles containing the matrix molecules 205 in which the molecular components of both the aerosol sample 201 and the matrix compound 204 are present. The matrix molecule 204 may be in excess of the molecular components of the aerosol sample 201.

The particles 205 may exit the sample transfer tube 202 and enter the inlet 206 of the ion analyzer 207. The particle 205 then enters the reduced pressure region 208, resulting in a substantial linear velocity due to the adiabatic expansion of the gas entering the vacuum region 208 from the sample transfer tube 202 and due to the associated free jet formation. get. The accelerated particles 205 may collide with the collision surface 209, and the collision event crushes the particles 205, eventually resulting in the formation of the molecular component gas phase ions 210 and the matrix molecules 211 of the aerosol sample 201. The collision surface 209 may be controlled and maintained at a temperature substantially higher than the ambient temperature.

The matrix 204 may contain a solvent for the sample 201 such that the sample 201 dissolves in the matrix 204, thereby removing the intramolecular adhesion between the sample molecules 201. Thus, when the lysed Specimen 205 then collides with the collision surface 209, the lysed Specimen 205 breaks into droplets, and every given droplet would have contained a specimen molecule that would otherwise have been contained in the matrix. Tends to contain fewer sample molecules. This makes the generation of sample ions 210 more efficient the next time the matrix in each droplet is evaporated. The matrix may contain organic solvents and / or volatile compounds. The matrix may include polar molecules, water, one or more alcohols, methanol, ethanol, isopanol, acetone, or acetonitrile. Isoprapanol is of particular interest.

The matrix molecule 211 may be freely diffused in vacuum. Conversely, the gas phase ion 210, which is a molecular component of the aerosol sample 201, may be transferred to the analysis region (not shown) of the ion analyzer 207 by the ion optics 212. The ions 210 may be guided to the analysis region by applying a voltage to the ion optics 212. The ions are then analyzed by the ion analyzer 207, which may include a mass spectrometer 102, an ion mobility analyzer, or a combination thereof. As a result of the analysis, chemical information about sample 201 may be obtained.

FIG. 3 shows a schematic view of an embodiment that is substantially similar to the embodiments shown and described with respect to FIG. However, sample 201 may be delivered by fluid / liquid transfer pump or Venturi pump 240 and matrix 204 may be delivered in liquid form. This allows the matrix compound 204 to be mixed into the aerosol 201 as water vapor or as a liquid before being introduced into the ion analyzer 207.

The Venturi pump 240 may include an inflow tube 242, which may be coupled to a device or probe (eg, a REIMS device or probe as described herein) and sample aerosol particles or liquids (eg, REIMS devices or probes as described herein). For example, it may be configured to transfer from biological tissue) to Venturi pump 240.

The Venturi pump 240 may include a gas inlet 244, which is a flow path for a liquid in which a gas (eg, nitrogen or standard medical air) is transferred to the Venturi pump 240 by aerosol particles 201 or inflow pipe 242. It may be arranged and adapted to be introduced into. The Venturi pump 240 may also include a discharge unit 246 for discharging the Venturi gas from the system so that the Venturi gas is not directed into the vacuum chamber 208 of the mass spectrometer 207.

The Venturi pump 240 may include a sample transfer section or capillary 202, and the sample transfer section or capillary 202 may be arranged and adapted to direct a mixture of sample and gas produced by the Venturi pump 240 toward contacts 248. Good. The matrix conduit 203 is arranged and adapted so that the matrix compound 204 is introduced into the contact 248 and the flow of the matrix compound 204 is directed to the inflow pipe 206.

The aerosol particles 201 and the matrix 204 may be internally mixed at the contacts 248, and the resulting aerosol particles 205 may be carried from the vacuum chamber 208 into the inflow tube 206 by exhalation. Larger aerosol particles 201 may not be carried into the inflow tube 206 because they are too heavy, and may pass through contacts 248 and exit the device via the discharge section 246.

Although shown as continuous in FIG. 3, the sample transfer section 202 may be a component separated from the contact point 248 and the inflow tube 206. The contact 248 may include a connector or connector (not shown) for connecting the individual sample transfer units 202. The connection between the contact 248 and the sample transfer section 206 may be fluid sealed and / or may include a ring clamp.

As described above, an important feature is the surface that induces dissociation of these clusters 205, following the formation of molecular clusters 205 containing the original specimen aerosol component 201 and matrix compound 204. The benefits of using Matrix 204 according to one embodiment can be seen in FIGS. 4A and 4B.

FIG. 4 shows the mass spectrum obtained by the sample undergoing the REIMS technique. With this REIMS technique, the aerosol was generated from the target, the aerosol collided with the heated collision surface, and the resulting ions were mass-analyzed. The mass spectrum of FIG. 4B was obtained by subjecting the same sample to the same analytical technique. However, the aerosol was mixed with the matrix (isoprapanol) before colliding with the collision surface and then mass spectrometrically analyzed. It can be seen from the two mass spectra of FIGS. 4A and 4B that the use of the matrix substantially increased the detected ionic strength.

It is believed that there are several mechanisms by which the addition of the matrix may improve the ionization of the specimen. For example, a mechanism may occur that results in protonation or deprotonation of the specimen. Alternatively or additionally, a reaction that removes water and / or ammonia from the specimen may occur. Alternatively or additionally, the addition of metal ions such as sodium may play a role in the mechanism. The most probable mechanism of protonation or deprotonation is similar to the MALDI method.

The dominant mechanism by which the addition of the matrix thereby enhances the ionization of the sample is believed to be by diluting or dissolving the sample so that ion dissociation promotes the formation of sample ions in the solution phase. Sample to be analyzed, Na +, ^{K +,} may contain counterions such as H ₃ O ^+, which interacts with the analyte to promote the formation of analyte ions in the solution phase. The resulting sample ions are then separated from the matrix in the gas phase when they collide with the surface (eg after collision with the collision surface and / or evaporation) via solventization, or the mechanism of the so-called lucky survivor of MALDI. You may.

A possible mechanism for enhancing the ionization matrix in ambient ionization techniques such as REIMS, depending on the matrix properties, is to form matrix (M) and sample (A) ions in solution as follows.

Alternatively, sample ionization may occur in the gas phase, as described, for example, in Biochimica et Biophysica Acta 1458 (2000) 6-27. However, this mechanism is considered to be less energetically preferable. The formation of matrix (M) and sample (A) ions in the gas phase may proceed as follows.

Ionization may occur in the gas phase or in the liquid phase. This carries the droplets in high concentration into the adjacent gas phase as the droplets containing the matrix and sample molecules dissolve rapidly upon contact with the collision surface.

Proton exchange may occur via two different types of translocation. Proton exchange may occur via an overbarrier or underbarrier (tunneling) transition, as there is a potential barrier to transfer. The probability of under-barrier transition depends on the shape of the barrier and the energy (E) of the tunneling particles. The energy dependence is very strong, almost a formal exp (E / ΔE) exponential, where ΔE is the energy characteristic for a given barrier. Explaining the probabilities of protons at energy level E, the probabilities of "pure" tunneling need to be multiplied by the Boltzmann factor exp (-E / kT). Therefore, the total contribution of level E is proportional to exp (E / ΔE−E / kT).

Therefore, there are two limiting cases. For ΔE >> kT, the probabilities tend to be exp (-E / kT), and the most probable mechanism is quantum mechanical behavior by tunneling from the ground state. Alternatively, for ΔE << kT, the most likely mechanism of transfer is the overbarrier transition, as the proton is most likely to be the highest possible level of E, the upper part of the barrier. In the first limiting case, the probability of proton transfer depends largely on the overlap of its vibrational wave functions, so the degree of improvement in ionization seen by the introduction of the matrix depends on the given sample and given matrix M. Become unique. This is seen, for example, in FIGS. 4A-4B, and it is clear that the improvement in ionization due to the addition of the matrix for ions with mass 766.6 Da is greater than the matrix for ions with mass 885.6 Da. is there.

Ｋｎｏｃｋｅｎｍｕｓｓ（Ａｎａｌｙｓｔ２００６、１３１ ９６６〜９８６）によれば、これは２つのステップ工程の最も物議を醸す側面が残っている。 Another possible mechanism is also similar to the MALDI mechanism and is a two-step process. The first step is the formation of the initial matrix (M) in the matrix sample solution as follows.

According to Knockenmus (Anallyst 2006, 131 966-986), this remains the most controversial aspect of the two-step process.

さらなる溶解により、クラスタ形成の場合に荷電分子イオンが分離してもよい。 The second step of the process may be involved in the ionic molecular reaction in the smoke that forms when the droplet hits the collision surface (which may or may not be heated) as follows.

Further dissolution may separate the charged molecular ions in the case of cluster formation.

FIG. 5A shows another embodiment of a mass spectrometer interface for introducing a sample aerosol and matrix into a mass spectrometer. This device comprises a Venturi pump 501. The Venturi pump 501 comprises a tube 502, which may be coupled to a device or probe (eg, a REIMS device or probe as described herein) to venturi the aerosol particles from a sample (eg, biological tissue). It may be configured to transfer to pump 501. The Venturi pump 501 may include a gas inlet 503, such that the gas inlet 503 introduces a gas (eg, Venturi gas) into the flow path of aerosol particles transferred into the Venturi pump 501 by a tube 502. It may be placed and adapted to. The Venturi pump 501 may include an enlarged sample transfer tube 504, which allows a mixture of sample and gas to flow from the tube 502 over the sampling device 510 via the outlet end 506 of the sample transfer tube 504. It may be arranged and adapted to be transferred to.

The sampling device 510 may broadly include a hollow tube or whistle 512, a matrix conduit 530, and an inflow tube 540. The matrix conduit 530 may be arranged and adapted to introduce the matrix in liquid form through channel 534 (FIG. 5B) within the matrix conduit 530. The matrix may leave the matrix conduit 530 through an end 534 placed or placed in the whistle 512, and the matrix may be sprayed by a gas attracted into the inflow tube 540. The quality of matrix atomization may be controlled and influenced by the dimensions and / or relative distances between the various parts of the sampling device 510, as described in more detail below.

The inflow tube 540 is connected to an ion analyzer or mass spectrometer, and a mixture of sample, gas and matrix passes through the end 542 of the inflow tube 540 placed or placed in the whistle 512, and the ion analyzer or mass. It may be arranged and adapted to pass through passage 544, which is transferred to the analyzer. In these arrangements, the collision surface 209 is arranged downstream of the inflow pipe 540.

FIG. 5C shows a proximity view of the sampling device 510.

The whistle 512 may be arranged to face the outlet end 506 of the sample transfer tube 504, a first side surface 522, and a second opposite that arbitrarily deviates from the outlet end 506 of the sample transfer tube 504. It may be provided in the form of a hollow tube with an arbitrary side surface 524.

The whistle 512 may include a first end 518, the first end 518 may be coaxially located about the inflow pipe 540, or may be hermetically engaged with it. The whistle 512 may include a second end 520, the second end 520 may be centered around a matrix conduit 530, or may be hermetically engaged with it.

A recess, opening, or notch 514 may be provided on the second side surface 524 of the whistle 512, the notch 514 with the sample flowing through the whistle 512 from the outlet end 506 of the sample transfer tube 504. An inlet may be formed so that the mixture of gases can be transferred inside the whistle 512.

A mixture of sample and gas exiting the outlet end 506 of the sample transfer tube 504 may collide with the first side surface 522 of the whistle 512 and then transfer around the outer surface and into the notch 514. .. Once the sample-gas mixture is inside the whistle 512, the sample-gas mixture is not arbitrarily transferred through the end 542 of the inflow tube 540 into the inflow tube 540. May be mixed with the mist-like matrix emerging from the matrix conduit 530. The sample, gas and matrix mixture may then be transferred to an ion analyzer or mass spectrometer via passage 544.

Positioning the notch 514 on the second side surface 524 of the whistle 512 means that the first collision of the sample-gas mixture is on a surface that is not directly exposed to the vacuum of the mass spectrometer. Thus, in various embodiments, the sampling device 510 is arranged and adapted such that the first collision of the mixture of sample and gas is on a surface that is not directly exposed to the vacuum of the mass spectrometer.

The notch 514 may have a substantially semicircular contour when the whistle 512 is seen in cross section (eg, as shown in FIGS. 5A and 5B). This means that the edge 517 of the notch 514 is oval when viewed from the direction facing the second side surface 524 of the whistle 512 (see FIG. 5C). Alternatively, the notch 514 may have a differently shaped contour when the whistle 512 appears to be, for example, a square, triangular, or irregularly shaped contour in cross section. The edge 517 of the notch 514 may also be square, triangular, or irregular when viewed from the direction in which the whistle 512 then faces the second side surface 524 of the whistle 12 (see FIG. 5C).

The position and orientation of the whistle 512 can affect the quantity and quality of the sample transferred into the mass spectrometer. The notch 514 may include a center point 516, which may coincide with the longitudinal centerline 508 of the sample transfer tube 504. FIG. 5C shows a view of the second side surface 524 of the whistle 512 (whistle 512 is shown separately in FIG. 5C), where the center point 516 can be seen as an oval center point.

The whistle 512 may be oriented such that the longitudinal axis 526 of the whistle 512 is aligned with the axis of symmetry of the notch 514. The center point 516 may be located on the axis of symmetry of the longitudinal axis 526 and / or the notch 514 of the whistle 512. The axis of symmetry of the notch 514 may include an axis of symmetry in the longitudinal direction, and the longitudinal direction may be defined as a direction along the longitudinal axis 526.

The location of various parts of the sampling device 510 can also affect the quantity and quality of samples transferred into the mass spectrometer.

Next, referring to FIG. 5B, the distance x is defined as the distance (eg, the shortest distance) between the end 534 of the matrix conduit 530 and the end 542 of the inflow pipe 540.

The distance y is defined as the distance (for example, the shortest distance) between the center point 516 of the notch 514 and the end 542 of the inflow pipe 540.

The distance z is defined as the distance (for example, the shortest distance) between the outlet end 506 of the sample transfer pipe 504 and the whistle 512 (for example, the first side surface 522 of the whistle 512).

The diameter a of the matrix conduit 530 can also affect the quantity and quality of the sample transferred into the mass spectrometer, affecting the atomization of the matrix as the matrix exits the end of the matrix conduit 530. be able to.

The diameter b of the inflow tube 540 and the diameter c of the sample transfer tube 504 can also affect the quantity and quality of the sample transferred into the mass spectrometer.

The diameters a, b, and c may correspond to the respective diameters of the end 532 of the matrix conduit 530, the end 542 of the inflow pipe 540, and the outlet end 506 of the sample transfer pipe 504.

All or all diameters a, b, and c are 0.2 mm, 0.4 mm, 0.6 mm, 0.8 mm, 1 mm, 1.2 mm, 1.4 mm, 1.6 mm, 1.8 mm, 2 mm, 2 mm, 2 .2mm, 2.4mm, 2.6mm, 2.8mm, 3mm, 3.2mm, 3,4mm, 3.6mm, 3.8mm, 4mm, 4.2mm, 4.4mm, 4.6mm, 4.8mm , Or may be longer, shorter, or substantially equal than 5 mm.

Any or all diameters / distances a, b, c, x, y, and z may be modified to optimize the quantity and quality of the sample transferred into the mass spectrometer.

Aspects of the present disclosure are the identification of one or more parameters associated with the sampling device 510, such as ion abundance or ion signal intensity, and the optimized value, maximum value, of the one or more parameters. Alternatively, it may extend to methods of optimizing the sampling device 510, including varying one or more distances a, b, c, x, y, and z to a minimum.

The Venturi pump 501 may be for introducing aerosol particles into the sample transfer tube 504. Sampling device 510 may be provided for sampling aerosols. The matrix conduit 530 may be arranged to introduce a matrix (such as isopropanol) into the sampling device 510, and the inflow tube 540 forwards a mixture of aerosol particles and matrix towards an ion analyzer or mass spectrometer. It may be arranged so as to face.

The Venturi pump 501 may facilitate the aspiration of aerosols or other gaseous samples containing the specimen, or may be driven by nitrogen or standard medical air. Aerosol sampling may be arranged to occur orthogonal to the outlet end 506 of the Venturi pump 501 as shown in FIGS. 1A and 1B. The outlet 532 of the matrix conduit 530 may be separated from the inflow tube 540 to the ion analyzer or mass spectrometer by a distance x. The distance x can be modified as needed to achieve the optimum ionic signal strength.

By changing the value of the distance x, the velocity of the gas attracted into the inflow pipe 540 can be changed, which can affect the atomization conditions. If the atomization conditions are less favorable, the matrix droplets do not have to be of the proper size to interact with the specimen aerosol and / or do not break effectively when the aerosol collides with the collision surface. You may.

FIG. 6 shows the ions acquired by the ion analyzer 207 for different distances x between the outlet 532 and the inlet 540 of the matrix conduit 530 when the matrix flow rate was set to about 0.2 ml / min. Indicates the signal strength. FIG. 6 shows ion signals for values of x = 0 mm, x = 1 mm, x = 2 mm, x = 4 mm, x = 4.5 mm, x = 5 mm, and x = 5.2 mm. It can be seen that when the distance x is about 0 mm (ie, the outlet of the matrix conduit 530 is in contact with the inlet 540), no ion signal is detected. When the distance x increases to about 1 mm, an ion signal is detected. As the distance x increases from about 1 mm to about 2 mm, the relative intensity of the ion signal increases. As the distance x increases from about 2 mm to about 4 mm, the relative intensity of the ion signal further increases. As the distance x further increases from about 4 mm to about 4.5 mm, the relative intensity of the ion signal decreases. As the distance x increases from about 4.5 mm to about 5 mm, the relative intensity of the ion signal is significantly reduced. When the distance x increases from about 5 mm to about 5.2 mm, the ion signal is virtually undetectable. This indicates that the detected ion signal can be optimized by selecting an appropriate value of x.

As the matrix 204 exits the matrix conduit 530, the matrix 204 may be atomized by a gas attracted into the ion analyzer inlet 240. By changing the value of the distance x, the velocity of the gas attracted into the ion analyzer inlet 240 changes, thus affecting the atomization conditions. If the atomization conditions are not preferred, the matrix droplets do not have to be of the proper size to interact with the specimen aerosol particles 201 and / or do not break sufficiently when colliding with the collision surface 209. You may.

The effect of different matrix 204 (eg isopropanol) flow rates on the appearance of the spectrum was tested.

FIG. 7 shows the ion analyzer 207 for different flow rates of the matrix 204 when the distance x between the outlet 232 of the matrix conduit 230 and the inlet 240 to the ion analyzer 207 is set to about 2.5 mm. Indicates the intensity of the ionic signal acquired by. The ion signal was measured at a flow rate of about 0.2 ml / min. The flow rate was then increased to about 0.4 ml / min and the intensity of the ionic signal increased. The flow rate was further increased to about 0.8 ml / min and the intensity of the ionic signal was reduced. The flow rate was then reduced to about 0.1 ml / min and the intensity of the ionic signal was reduced. The flow rate was then further reduced to about 0.05 ml / min and the intensity of the ionic signal was further reduced. The flow rate was then further reduced to about 0.025 ml / min and the intensity of the ionic signal was further reduced. The flow rate was then further reduced to about 0.01 ml / min and the intensity of the ionic signal was further reduced. This indicates that the ion signal does not have to be simply improved by increasing the flow rate of the matrix 204, but the flow rate may be optimized to produce the optimum ion signal intensity.

8A-8I show the effect of different isoprapanol flow rates on the REIMS spectral contours for Bacteroides fragilis using flow rates of 0.01-0.25 ml / min. 8A-8I show spectra for a mass range of 500-900 covering the phospholipid sample. The effect of isoprapanol present in the analysis of this sample is detectable from a very low rate, eg 0.02 ml / min, the appearance of m / z 590 (ceramide species) and m / z 752 (α galactosylceramide) in the spectrum. Seen clearly from. These species were found to increase in their relative abundance as the flow rate of isoprapanol increased further.

Although it has been described in FIG. 5 that the matrix is introduced downstream of the sample transfer tube 504, as opposed to the inlet 240 to the ion analyzer 207, the matrix is optionally introduced into the sample transfer tube 504. You may.

Alternatively, the matrix may be introduced at a location around the transfer tube 504 or may be swept into the inlet 240 by airflow towards the inlet 240 to the ion analyzer 207.

Calibration / Rockmass / Lock Mobility compounds may be used in the various techniques described herein to calibrate the ion analyzer or provide a reference mass to the ion analyzer. The calibration, rock mass, or lock mobility compounds may be introduced via the matrix conduit 203, via the sample transfer tube 202, or elsewhere.

9A and 9B show two mass spectra obtained by analyzing a sample of porcine muscle according to one embodiment. While the spectrum of FIG. 9A was acquired, the rockmas compound (leucine enkephalin) was introduced into the analyzer 207 through the matrix conduit 203. The peak for the rock mass compound can be observed as the first peak in the mass spectrum. The mass-to-charge ratio of rock mass ions is known in advance and can be used to calibrate the mass spectrometer 207 so that the mass-to-charge ratio of other detected ions can be determined more accurately. The mass spectrum shown in FIG. 9B was obtained using the same method as the spectrum in FIG. 9A. However, no rock mass compound was used in the analysis. It can be seen that the two mass spectra are substantially identical, except that rock mass ions were detected in the mass spectra of FIG. 9A. Therefore, it is clear that the introduction of the rock mass compound in this technique does not affect the mass spectrum measured by the ion analyzer 207.

FIG. 10 shows a graph obtained from principal component analysis of porcine brain over 4 days. Data for days 1 and 4 were obtained without the rockmas compound, whereas data for days 2 and 3 were obtained using the rockmas compound. The rock mass ions are not shown in the graph because the analysis was performed over a mass unit range of 600-900 and therefore rock mass ions are outside this range (see FIG. 9A). Principal component analysis does not distinguish the data obtained with the Rockmass compound from the data obtained without the Rockmass compound, and the differences due to the data from different dates are Rockmass. Shows that it is significantly greater than any difference due to the inclusion of compounds.

FIG. 11 shows the results of principal component analysis for analysis of data obtained from porcine liver cortex, porcine liver, porcine brain, porcine myocardium, and other porcine muscles. Some data were obtained with the Rockmass compound and some were obtained without the Rockmass compound. However, data for all types of tissues are well clustered within specific regions of the graph, demonstrating that the use of rockmas compounds does not affect analysis and tissue classification.

It was determined that two or more different known rockmas compounds could be used without adversely affecting the analysis of the sample. Exactly the rock mass compound (s) may be used and / or the external rock mass compound (s) may be used.

12A and 11B show schematic configurations of exemplary configurations of collision surfaces that may be used in the present invention. FIG. 12A corresponds to the collision surface 209 shown in FIGS. 2 and 3. For example, the collision surface 209 may be a spherical stainless steel collision surface 209a, and may be mounted in the analyzer 207 from the end of the inflow capillary 206 by about 6 mm. FIG. 12B shows an alternative collision surface 209 that may be used in the form of a coil-shaped collision surface 209b. Ions may be transferred by ion optics 212 to the analytical region (not shown) of ion analyzer 207. As discussed above, the ion optics 212 may include a Stepwave® ion guide.

It is contemplated that the collision surface may be substantially other shapes such as cylindrical, tubular, rod, hemispherical, teardrop, disk, concave, dish, or conical. It is also contemplated that the collision surface may be formed by the inner surface of a hollow collision assembly having an inlet and an outlet. Aerosols may enter through the inlet and then slam into the inner surface of the collision assembly to form or release sample ions. Specimen ions may then emerge from the collision assembly via the outlet. The internal cross-sectional area of the collision assembly may be substantially universal or reduced in the direction from the inlet to the outlet, ie the collision assembly may be funnel-shaped, tubular-shaped, or cylindrical. .. Embodiments relating to hollow funnel-shaped collision assemblies or hollow cylindrical collision assemblies have also been found to result in high ion yields (or improved ionization efficiencies) coupled with significant improvements in the signal-to-noise ratio. It was also found that these embodiments are less likely to result in collision assembly and downstream ion optics contamination due to background clusters of non-analytical interest.

The REIMS mechanism recognizes that the generation of positively and negatively charged ions can occur substantially equally, with positively and negatively charged ions subsequently being molecular clusters with relatively large neutral charges. May form. These neutral clusters are not very well manipulated by the electric field in the analyzer or analyzer and may therefore be removed, for example, by instrumental ion optics 212. The collision plane 209 described herein serves to split the molecular cluster 205 and release the ions, so that the ions may be guided by an electric field in the analyzer or analyzer. However, it is also recognized that the provision of collision surface 209 can induce secondary contamination between measurement results of different samples. For example, lipopolypeptides such as certain bacterial metabolites, such as certain sphingolipids produced by the genus Bacillus or surfactins and lichenicins produced by the genus Bacillus, have undergone only a few repeated measurements It was found to induce a relatively strong memory effect. This secondary contamination can be mitigated by cleaning the atmospheric interface prior to each analysis. However, this is not particularly desirable for automatic appliances. The surface may be heated to, for example, several hundred degrees Celsius to prevent contamination of the collision surface 209. For example, by heating the collision surface 209, carbonaceous deposits on the collision surface 209 may react with oxygen introduced through the inflow capillaries 206. Then the carbonaceous deposits is converted to CO ₂ gas, CO ₂ gas can be separated from the impact surface 209, does not contaminate the instrument during hence subsequent analysis. The coil-shaped collision surface 209b of FIG. 12B provides particularly reproducible heat distribution.

The collision element or collision surface 209 may be composed of a material that may be heated by passing an electric current through the collision surface 209, for example by applying a voltage V in FIG. 12B, and the collision surface during analysis. Can be easily heated. For example, the collision surface 209 may be manufactured from a heat resistant iron chromium aluminum (FeCrAl) alloy such as Kanthal. By using such a heated collision surface 209, the memory effect may be significantly reduced and therefore the frequency of cleaning the instrument may be significantly reduced. For example, thousands of database entries could be recorded without any amnestics, and no carry-over was seen with extended exposure to lipopolypeptides.

The spectral profile acquired using the heated collision surface 209 may differ from the spectral profile acquired using the unheated collision surface 209, for example as shown in FIGS. 13A and 13B. There is.

13A and 13B show spectral profiles resulting from the analysis of Bacteroides fragilis using the unheated and heated collision surfaces, respectively. This indicates that not all of the spectral components analyzed using this type of heated surface technique are sufficiently thermally stable. For example, the effect of the heating surface seems to be particularly strong against phosphatidic acid (which is common in fungi such as Candida albicans) and sphingolipids (which are common in Bacteroides), while phosphatidylglycerols. And phosphatidylethanolamine (these are, for example, the major phospholipid species in Mirabilis deforming fungi) have generally less effect on spectral appearance.

As explained above, it has been found that the introduction of matrix compound 204, such as isopropyl alcohol (IPA), upstream of the collision surface 209 improves specimen ionization and instrument sensitivity. It has also been found that the introduction of matrix compound 204 may restore spectral properties that would otherwise be impaired by the use of heated collision surfaces rather than unheated collision surfaces. For example, FIGS. 13A and 13B demonstrate that the use of heated collision surfaces has been found to eliminate spectral properties such as ceramides within Bacteroides fragilis. By introducing isoprpanol into the sample aerosol 201 prior to introduction into the mass spectrometer 207 or analyzer, these spectral characteristics are restored, similar to the fingerprint of an atmospheric interface with an unheated collision surface. It was found to produce a fingerprint of the mass spectrum of. In addition, the addition of matrix 204 (eg, isoprpanol) to the sample aerosol 201 resulted in similar or higher signal strength compared to direct introduction of the aerosol, allowing the Venturi pump 213 to be used for aerosol transport.

14A-14C show three mass spectra obtained by analyzing a sample of Candida albicans (yeast). The mass spectrum of FIG. 14A was obtained while introducing the isopropyl alcohol (IPA) matrix upstream of the heated collision surface according to one embodiment. The mass spectrum of FIG. 14B was obtained using the same method as the spectrum of FIG. 14A, except that a matrix without isopropyl alcohol (IPA) was introduced. The mass spectrum of FIG. 14C was obtained using the same method as the spectrum of FIG. 14B, except that the collision surface was not heated. 14A-14C show the effect of the heated surface and the isoprapanol matrix on the spectral appearance of Candida albicans (yeast). In these examples, the use of heated collision surfaces instead of unheated collision surfaces significantly alters the spectral appearance, and many spectral properties within Candida albicans significantly reduce or completely eliminate relative intensities. Show that. Introducing isoprapanol into a system with a heated collision surface helps to avoid this problem and produces a more similar spectrum to the spectrum obtained using the unheated collision surface.

As described above, rock mass compounds (leucine, enkephalin, etc.) may be used. It is herein that the peak intensity of the rock mass compound is monitored and may be used to determine if the matrix 204 flows at the desired rate, eg, by introducing the rock mass compound along the matrix 204. Intended in the book. This may be used to determine that matrix compound 204 flows consistently and is not at a variable flow rate.

In order to calibrate the instrument described herein, a calibrator may be introduced into the instrument and analyzed. For example, the instrument may be calibrated prior to analysis of the sample (eg, tissue) and then determined for any mass shift. The calibrator (eg, sodium formate) may be injected into the instrument using a matrix (eg, isoprapanol) injection tube 203. However, the optimum distance between the outlet 232 of the matrix injection tube 230 and the inlet 240 of the mass spectrometer 207 or the analyzer for calibration may be shorter than the optimum distance x for sample analysis, eg, microstructure analysis. It's been found. To address this, different IPA capillary lengths may be used during calibration and tissue analysis. Relatively long capillaries may be used for calibration, whereas shorter capillaries may be used for tissue analysis.

As described above, the matrix conduits may be arranged in a variety of different configurations. For example, the matrix conduit may be coaxial with the inflow pipe to the mass spectrometer or may be located inside the inflow pipe of the mass spectrometer. The distance from the outlet of the matrix conduit to the downstream outlet of the inflow tube of the mass spectrometer (ie, the inlet to the vacuum chamber) was found to be important. It is thought that the longer the distance, the better the interaction between the sample and the matrix.

FIG. 15A shows the total ion current detected as a function of time for several different distances between the outlet of the matrix conduit and the inlet to the mass spectrometer vacuum chamber. A positive distance represents the downstream distance of the inlet of the mass spectrometer to the vacuum chamber, while a negative distance represents the upstream distance of the inlet of the mass spectrometer to the vacuum chamber. The sample analyzed was porcine liver and the matrix was isopropyl alcohol. The matrix capillaries were made of quartz glass and had an outer diameter of 360 μm and an inner diameter of 250 μm. Relatively strong background noise and strong IPA signals were observed when the outlet of the matrix conduit was located upstream of the inlet to the vacuum chamber of the mass spectrometer (ie, negative distance).

15B-15F show mass spectra acquired at different distances in FIG. 15A. 15B-15F show mass spectra obtained at distances of -10 mm, -20 mm, -24 mm, 0 mm, and + 2 mm, respectively. It can be seen that the effect of the matrix was relatively low when the outlet of the matrix conduit was located at the inlet to the vacuum chamber or downstream of the inlet (ie 0 mm and +2 mm distance). Conversely, the greater the matrix conduit outlet was located upstream of the inlet to the vacuum chamber (ie, the greater negative distance), the greater the effect of the matrix. It was confirmed that increasing the flow rate of the matrix at a distance of 0 mm did not improve the observed total ion current.

FIG. 16 shows the data corresponding to the data in FIG. 15A, except that the data were acquired using a matrix conduit having an inner diameter of 100 μm and a different distance to FIG. 15A was used. Similar to FIG. 15A, the observed ionic signal increased as the outlet of the matrix conduit was located more upstream of the inlet to the vacuum chamber (ie the distance was greater negative).

FIG. 16B shows mass spectra acquired at different distances from FIG. 16A. 16B-16F show mass spectra obtained at distances of 0 mm, +2 mm, -1 mm, -10 mm, -20 mm, -30 mm, -40 mm, and -50 mm, respectively. It can be seen that the effect of the matrix was relatively low when the outlet of the matrix conduit was located at the inlet to the vacuum chamber or downstream of the inlet (ie 0 mm and 2 mm distances). Conversely, the greater the matrix conduit outlet was located upstream of the inlet to the vacuum chamber (ie, the greater negative distance), the greater the effect of the matrix. The spectrum was less noisy than the spectra of FIGS. 15B to 15F, and the matrix had a stronger effect. It was confirmed that increasing the flow rate of the matrix at a distance of 0 mm did not improve the spectrum.

FIG. 17A shows the data corresponding to the data of FIG. 15A, except that the data was acquired using a matrix conduit having an inner diameter of 50 μm and a different distance to FIG. 15A was used. Similar to FIG. 15A, the ionic signal observed in FIG. 17A increased as the outlet of the matrix conduit was located more upstream of the inlet to the vacuum chamber (ie the distance was greater negative).

17B to 17IB show mass spectra acquired at different distances from FIG. 17A. 17B-17F show mass spectra obtained at distances of 0 mm, +2 mm, -2 mm, -10 mm, -20 mm, -30 mm, -40 mm, and -50 mm, respectively. It can be seen that the effect of the matrix was minimal when the outlet of the matrix conduit was located at the inlet to the vacuum chamber or downstream of the inlet (ie 0 mm and 2 mm distances). Conversely, the greater the matrix conduit outlet was located upstream of the inlet to the vacuum chamber (ie, the greater negative distance), the greater the effect of the matrix.

It was found that the position of the matrix conduit has a greater effect on the ionic signal than the flow rate of the matrix.

18A-18C show that the outlets of each matrix conduit are located 20 mm upstream of the inlet to the vacuum chamber and have inner diameters of 50 μm, 100 μm, and 250 μm, respectively, when using a matrix flow rate of 0.2 ml / min. The three spectra obtained for the matrix conduit are shown. The spectrum shows that the smaller the inner diameter of the matrix conduit, the better the spectrum and the less noise.

It was also found that tapering the outlet end of the matrix conduit improved the detected ionic signal intensity.

As described above, whistle arrangements may be used for sampling, for example with respect to FIG. 5B. In this arrangement, the matrix conduit may be coaxial with the inflow tube to the mass spectrometer. As explained above, it has been found that the distance x from the outlet of the matrix conduit to the inlet of the mass spectrometer is important.

FIG. 19A shows the total ion current detected as a function of several different distances between the outlet of the matrix conduit and the inlet of the mass spectrometer to the inflow tube in the whistle arrangement. The sample analyzed was porcine liver and the matrix was isopropyl alcohol. The matrix capillaries were made of quartz glass and had an outer diameter of 360 μm and an inner diameter of 250 μm. It can be seen that the ionic signal intensities were about the same for different distances between the outlet of the matrix conduit and the inlet of the mass spectrometer, up to a distance of about 3-4 mm.

19B-19H show mass spectra acquired at different distances from FIG. 19A. 19B-19F show mass spectra obtained at distances of 6 mm, 5 mm, 4 mm, 3 mm, 2 mm, 1 mm, and 0 mm, respectively. It can be seen that the spectra are very similar at distances up to about 3 mm.

FIG. 20A shows the data corresponding to the data of FIG. 19A, except that the data was acquired using a matrix conduit having an inner diameter of 100 μm. It can be seen that the ionic signal intensity is about the same up to about 3 mm (although the highest intensity at a distance of about 2 mm) for different distances between the outlet of the matrix conduit and the inlet of the mass spectrometer. .. The ionic signal intensity dropped at distances over 3 mm.

20B-20G show mass spectra acquired at different distances from FIG. 20A. 20B-20G show mass spectra obtained at distances of 5 mm, 4 mm, 3 mm, 2 mm, 1 mm, and 0 mm, respectively.

FIG. 21A shows the data corresponding to the data of FIG. 19A, except that the data was acquired using a matrix conduit having an inner diameter of 50 μm and the data for additional distances are shown. It can be seen that the ionic signal intensities are about the same up to about 4 mm for different distances between the outlet of the matrix conduit and the inlet of the mass spectrometer. Ion signal intensity dropped at distances greater than 4 mm.

21B-21I show mass spectra acquired at different distances from FIG. 21A. 21B-21I show mass spectra obtained at distances of 7 mm, 6 mm, 5 mm, 4 mm, 3 mm, 2 mm, 1 mm, and 0 mm, respectively.

Various biological samples were analyzed using methods and techniques according to various embodiments of the invention. These analyzes demonstrated that the use of collision planes and / or matrices improved the ionic signals obtained from the specimens.

Each of FIGS. 22A-22C shows mass spectra obtained using the REIMS technique in negative ion mode for analysis of lamb liver. Each spectrum represents data obtained from 5 samples. The spectrum of FIG. 22A was acquired without introducing the matrix into the sample stream and without using the collision plane. The spectrum of FIG. 22B was obtained by introducing a matrix (isopropyl alcohol at a rate of 0.2 mL / min) into the sample stream and using no collision plane. The spectrum of FIG. 22C was obtained using a collision plane with a matrix (isopropyl alcohol at a rate of 0.2 mL / min) introduced into the sample stream. By comparing these spectra, the signal intensity of the sample ions is increased by using the matrix without using the collision surface, and the signal intensity of the sample ions is increased by using the matrix and the collision surface in combination. It can be seen that there is a significant increase.

Each of FIGS. 23A-23C shows mass spectra obtained using the REIMS technique in positive ion mode for analysis of lamb liver. Each spectrum represents data obtained from 5 samples. The spectrum of FIG. 23A was acquired without introducing the matrix into the sample stream and without using the collision plane. The spectrum of FIG. 23B was obtained by introducing a matrix (isopropyl alcohol at a rate of 0.2 mL / min) into the sample stream and using no collision plane. The spectrum of FIG. 23C was obtained by introducing a matrix (isopropyl alcohol at a rate of 0.2 mL / min) into the sample stream and using the collision plane. Similar to the negative ion modes shown in FIGS. 22A to 22C, by comparing the positive ion modes of FIGS. 23A to 23C, the signal intensities of the sample ions can be obtained by using the matrix without using the collision surface. It can be seen that the signal strength of the sample ions is significantly increased by using the matrix and the collision surface in combination.

It has also been found that analysis of high adipose tissue, such as normal breast tissue, may generate little or no ionic signal without the use of a matrix.

FIG. 24A shows mass spectra obtained from analysis of normal breast tissue without the use of a matrix. FIG. 24B shows the mass spectrum obtained from the analysis of normal breast tissue using isopropyl alcohol as a matrix. By comparing these spectra, it can be seen that the use of the matrix significantly increases the signal intensity of the sample ions.

Analysis of Sample Spectrum The table below provides a list of analytical techniques intended to fall within the scope of the present invention.

The above-mentioned analysis methods such as PCA-LDA, PCA-MMC, and PLS-LDA can be used in combination.

By analyzing the sample spectrum, an analysis without an objective variable for dimensional degeneracy can be followed by an analysis with an objective variable for classification.

As an example, a number of different analytical techniques will be described in more detail.

Multivariate analysis to develop a model for classification Next, as an example, a method of constructing a classification model using multivariate analysis of multiple standard sample spectra will be described.

FIG. 25 shows method 1500 for constructing a classification model using multivariate analysis. In this example, the method comprises step 1502 to obtain multiple sets of intensity values relative to the standard sample spectrum. The method then includes step 1504 of principal component analysis (PCA) without objective variables, followed by step 1506 of linear discriminant analysis (LDA) with objective variables. This technique is sometimes referred to herein as PCA-LDA. Other multivariate analysis techniques such as PCA-MMC may be used. The PCA-LDA model is then output in step 1508 for storage, for example.

Multivariate analysis, such as this step, provides a classification model in which an aerosol, smoke, or vapor sample can be classified using one or more sample spectra obtained from an aerosol, smoke, or vapor sample. can do. Next, multivariate analysis will be described in more detail in relation to a simple example.

FIG. 26 shows a set of standard sample spectra obtained from two classes of known standard samples. The class may be any one or more classes of targets described herein. However, for the sake of clarity, we will refer to the two classes as the left class and the right class in this example.

Each standard sample spectrum is preprocessed to derive a set of three standard peak intensity values for each mass-to-charge ratio in terms of the standard sample spectrum. Only three standard peak intensity values are shown, but more standard peak intensity values (eg, about 100 standard peak intensity values) are for the corresponding number of mass-to-charge ratios in each standard sample spectrum. It will be understood that it may be pulled out. In other embodiments, the standard peak intensity values may correspond to mass, mass-to-charge ratio, ion mobility (drift time), and / or operating parameters.

FIG. 27 shows a multivariate space with three dimensions defined by the intensity axis. Each dimension or intensity axis corresponds to the peak intensity, especially at the mass-to-charge ratio. Again, it will be appreciated that there may be more dimensions or intensity axes (eg, about 100 dimensions or intensity axes) in a multivariate space. The multivariate space comprises multiple reference points, each reference point corresponding to a standard sample spectrum, i.e., the peak intensity value of each standard sample spectrum provides coordinates with respect to a reference point in the multivariate space.

A set of standard sample spectra is a matrix element that is the row associated with each standard sample spectrum, the column associated with each mass-to-charge ratio, and the peak intensity value for each mass-to-charge ratio of each standard sample spectrum. It may be represented by the reference matrix D having.

In many cases, the large number of dimensions and matrix D in a multivariate space can make it difficult to group standard sample spectra into classes. The PCA may perform Matrix D accordingly to calculate a PCA model that defines the PCA space with a reduced number of one or more dimensions defined by the principal component axis. The principal component may be selected to include or "explain" the maximum multivariate space in matrix D, or to be a principal component that cumulatively describes the threshold of the maximum multivariate space in matrix D.

FIG. 28 shows how the cumulative multivariate can be increased as a function of the number n of principal components in the PCA model. Multivariate thresholds may be selected as desired.

The PCA model may be calculated from matrix D using the nonlinear iterative least squares regression (NIPALS) algorithm or singular value decomposition, and details of the nonlinear iterative least squares regression (NIPALS) algorithm or singular value decomposition are known to those skilled in the art. Therefore, it will not be described in detail in this specification. Other methods of calculating the PCA model may be used.

The resulting PCA model may be defined by a PCA scoring the matrix S and a PCA deriving the matrix L. The PCA may also generate an error matrix E, which contains multivariates not explained by the PCA model. The relationship between D, S, L, and E may be as follows.
^{D = SL T + E (1} )

FIG. 29 shows the PCA space obtained for the standard sample spectra of FIGS. 26 and 27. In this example, the PCA model has two principal components PC ₀ and PC ₁ , so the PCA space has two dimensions defined by two principal component axes. However, fewer or more principal components may be included in the PCA model as desired. It is generally desirable that the number of principal components be at least one less than the number of dimensions in the multivariate space.

The PCA space contains a plurality of transformed reference points or PCA scores, and each transformed reference point or PCA score corresponds to the standard sample spectrum of FIG. 26 and thus corresponds to the reference point of FIG. 27. ..

As shown in FIG. 29, the reduced dimensions of the PCA space make it easier to group standard sample spectra into two classes. Any outliers may also be identified and removed from the classification model at this stage.

Even if a multivariate analysis with more objective variables is performed, such as a multiclass LDA in PCA space or a Principal Component Component Analysis (MMC), to define classes and arbitrarily further reduce dimensions. Good.

As will be appreciated by those skilled in the art, multiclass LDAs should maximize the ratio of variance between classes to variance within classes (ie, give the maximum possible distance between the smallest possible classes). )try to. The details of LDA are known to those skilled in the art and will not be described in detail herein.

The resulting PCA-LDA model may be defined by a transformation matrix U, which solves the generalized eigenvalue problem for each of the transformed spectra contained therein. It may be derived from the PCA score matrix S and class assignments.

The conversion of the score S from the original PCA space to the new LDA space may then be given by:
Z = SU (2)
In the above equation, the matrix Z contains the score converted into the LDA space.

FIG. 30 shows a PCA-LDA space with one dimension or axis, and the LDA is performed within the PCA space of FIG. As shown in FIG. 30, the LDA space contains a plurality of further converted reference points or PCA-LDA scores, and each further converted reference point becomes the converted reference point or the PCA score of FIG. 29. Correspond.

In this example, the dimensions of the PCA-LDA space are further reduced, making it easier to group the standard sample spectra into two classes. Each class in the PCA-LDA model is an average and covariance matrix of the transformed class or one or more hyperplanes (including points, lines, planes, or higher dimensional hyperplanes) or hypersurfaces or It may be defined by a boronoy cell in the PCA-LDA space.

The PCA loading matrix L, the LDA matrix U, and the transformed class mean and covariance matrix or hyperplane or hypersurface or boronoy cells are databases for later use in classifying aerosol, smoke, or water vapor samples. It may be output to.

The transformed covariance matrix in the LDA space V'g for class g may be given by:
_V'g = V ^T V _g U (3)
In the above equation, V _g is a class covariance matrix in PCA space.

Average position Z _g of the converted class for class g may be given by the following.
S _g U = Z _g (4)
In the above equation, S _g is the average position of the class in the PCA space.

Multivariate analysis using a model for classification The following example describes how to use classification to classify aerosol, smoke, or vapor samples.

FIG. 31 shows a method 2100 using a classification model. In this example, the method includes step 2102 to obtain a set of intensity values for the sample spectrum. The method then comprises projecting a set of intensity values for the sample spectrum into the PCA-LDA model space in step 2104. Other classification model spaces such as PCA-MMC may be used. The sample spectrum is then classified in step 2106 based on the projected position, and then in step 2108 the classification is output.

The classification of aerosol, smoke, or vapor samples will then be described in more detail with respect to the simple PCA-LDA model described above.

FIG. 32 shows a sample spectrum obtained from an unknown aerosol, smoke, or vapor sample. The sample spectra are preprocessed to derive a set of three sample peak intensity values for each mass-to-charge ratio. As mentioned above, only three sample peak intensity values are shown, but more sample peak intensity values (eg, about 100 sample peak intensities) at more corresponding mass-to-charge ratios to the sample spectrum. It will be understood that the value) may be derived. Also, as described above, in other embodiments, the sample peak intensity values may correspond to mass, mass-to-charge ratio, ion mobility (drift time), and / or operating parameters.

Sample spectrum may be represented by the sample vector d _x, the elements of the vector is the peak intensity values for each mass-to-charge ratio. The PCA vector s _x converted with respect to the sample spectrum can be obtained as follows.
d _x L = s _x (5)

The PCA-LDA vector z _x then transformed with respect to the sample spectrum can then be obtained as follows.
s _x U = z _x (6)

FIG. 33 also shows the PCA-LDA space of FIG. However, the PCA-LDA space of FIG. 33 further includes projected sample points corresponding to the transformed PCA-LDA vector z _x derived from the peak intensity values of the sample spectrum of FIG. 32.

In this example, the projected sample point is for one side of the hyperplane between the classes for the right class, so the aerosol, smoke, or vapor sample may be classified to belong to the right class.

Alternatively, may be used Mahalanobis distance from the class center in LDA space, Mahalanobis distance between the point z _x from the center of this case class g may be given by the following square root,
^{_{(Z x -z g) T (}} V 'g) -1 (z x -z g) (8)
Data vector d _x is the distance may be assigned to a class which is the minimum.

In addition, processing each class as a multivariate gauss may calculate the probabilities of the members of the data vector for each class.

Library-Based Analysis, Developing a Library for Classification Next, as an example, we describe how to build a classification library using multiple input standard sample spectra.

FIG. 34 shows a method 2400 for constructing a classification library. In this example, the method comprises acquiring multiple input standard sample spectra and step 2404 to extract metadata from the plurality of input standard sample spectra for each class of sample. The method then includes step 2406 storing metadata for each class of sample as a separate library registration. The classification library is then output in step 2408, for example, to an electronic storage device.

Such a classification library can classify aerosol, smoke, or vapor samples using one or more sample spectra obtained from aerosol, smoke, or vapor samples. Next, the library-based analysis will be described in more detail with reference to an example.

In this example, each registration in the classification library is generated from multiple preprocessed standard sample spectra that represent the class. In this example, the standard sample spectrum for the class is preprocessed according to the following procedure.

上式で、Ｎ_ｃｈａｎは選択された値であり、

は、ｘより下の最も近い整数を表す。一例では、Ｎ_ｃｈａｎは２^１２すなわち４０９６である。 First, the rebinning process is executed. In this embodiment, the data is resampled on a logarithmic grid in abscissa.

In the above equation, N _chan is the selected value,

Represents the closest integer below x. In one _{example, N chan} is ^{2 12} i.e. 4096.

Then the background subtraction process is executed. In this embodiment, a cubic spline with k knots is then constructed such that the p% of the data between each pair of knots is below the curve. This curve is then subtracted from the data. In one example, k is 32. In one example, p is 5. Then, the constant value corresponding to the q% quantile value of the data obtained by subtracting the intensity is subtracted from each intensity. Positive and negative values are maintained. In one example, q is 45.

の平均値を有するように正規化される。一例では、

である。 Normalization is then performed. In this embodiment, the data is

Normalized to have an average value of. In one example

Is.

Then registration of the library consists of the form of metadata in the central spectral values mu _i and deviation value D _i for each N _chan point in the spectrum.

上式で、１／２≦Ｃ＜∞であり、Ｉ’（Ｃ）はガンマ関数である。 The likelihood for the i-th channel is given by:

In the above equation, 1/2 ≤ C <∞, and I'(C) is a gamma function.

The above equation is a generalized Cauchy distribution that is reduced to a standard Cauchy distribution for C = 1 and becomes a Gaussian (standard) distribution as C → ∞. The parameter D _i controls the width of the distribution (in the Gaussian limit, _Di = σ _i is simply the standard deviation), while the global value C controls the size of the tail.

In one example, C is 3/2, which is between Cauchy and Gauss, so the likelihood is:

For each library registration, while the parameter mu _i is set to the median value of the list of values in the channel of the i input standard sample spectrum, quarter of these values divided by the difference D _i is √2 Taken in the interquartile range. This selection ensures that the likelihood for the i-th channel has the same interquartile range as the input data, and the use of the interquartile provides some protection from out-of-range data.

Library-Based Analysis Using the Library for Classification The following example describes how to use the classification library to classify aerosol, smoke, or vapor samples.

FIG. 35 shows a method 2500 using a classification library. In this example, the method comprises step 2502 to acquire a set of multiple sample spectra. The method then includes step 2504 to calculate the probabilities or classification scores for a set of multiple sample spectra for each class of samples using metadata for class registration in the classification library. Then, the sample spectrum is classified in step 2506, and then the classification is output in step 2508.

The classification of aerosol, smoke, or vapor samples will then be described in more detail with reference to the classification library described above.

In this example, the unknown sample spectrum y is the median spectrum of a plurality of sample spectra in a set. By taking the median spectrum y, it is possible to protect from out-of-range data for each channel.

上式で、μ_ｉおよびＤ_ｉは、それぞれチャネルｉに対するライブラリ中央値および偏差値である。尤度Ｌ_ｓは数的安全性に対するログ尤度として計算されてもよい。 Then the likelihood L _s for the input data provided in the registration of the library is provided by.

In the above equation, μ _i and _Di are the library median and deviation values for channel i, respectively. Likelihood L _s may be calculated as the log likelihood for numerical safety.

に対して得られる確率が以下によって与えられる。

The likelihood L _s is then normalized over the S'of all candidate classes to give the probabilities, assuming that the prior probabilities are uniform across the classes. class

The probabilities obtained for are given by:

The exponent (1 / F) can mitigate the probability that it might otherwise be too limited. In one example, F = 100. These probabilities may be expressed as percentages, for example in the user interface.

Alternatively, the RMS classification score R _s may be calculated using the same central sample values as the library and derivative values from the library.

Again, the score Rs is normalized over the S'of all candidate classes. Aerosol, smoke, or vapor samples may then be classified as belonging to the class with the highest probability and / or highest RMS classification score.

Methods of treatment, surgery and diagnosis, as well as non-medical methods A variety of different embodiments are contemplated. According to some embodiments, the methods disclosed above may be performed on tissues inside the body, tissues outside the body, or tissues in vitro. Tissue may include human or non-human animal tissue.

Various surgical methods, treatment methods, medical methods, and diagnostic methods are planned.

However, it is intended to be related to non-surgical and non-therapeutic methods of mass spectrometers that are not performed on tissues in the body. Other related embodiments are intended to be performed in an extracorporeal manner, such that those embodiments are performed outside the human or animal body.

A further embodiment is intended in which the method is performed on a non-surviving human or animal, eg, as part of an autopsy procedure.

The various embodiments described herein provide an instrument and related methods for the chemical analysis of aerosol and gaseous samples containing specimens using mass spectrometers or other vapor phase ion analysis modalities. .. The method begins by introducing a sample of aerosol or other gas containing the sample 201 into the enclosed space. In the enclosed space, sample 201 is mixed with low molecular weight matrix compound 204. This homogeneous or heterogeneous mixture is then introduced into the atmospheric interface of the mass spectrometer 102 or ion mobility analyzer via the inlet 206. When the mixture is introduced into the low pressure region of the analyzer, aerosol particles containing the molecular components of the sample and matrix compounds are formed and the aerosol particles are accelerated by the expansion of the free jet. The mixed composition aerosol particles 205 are subsequently dissolved through collisions with the solid collision surface 209. The dissolution event produces neutral and charged species containing the molecular ions 210 of the chemical constituents of the sample. Ion 210 is separated from the neutral species by using an electric field to guide the ions 210 on different pathways to the neutral species, for example by using an ion guide 212 such as Stepwave® ion guide. May be done. The molecular ion 210 is then subjected to mass or mobility analysis. This provides a simple solution for the analysis of the molecular components of the aerosol online, without the application of high pressure or laser.

This method and device provides solutions for online mass spectrometry and ion mobility spectroscopy of vapor or aerosol samples.

According to various embodiments, the matrix compound 204 may be mixed into the sample aerosol 201 at all points, either as a vapor or as a liquid, before introducing the sample into the ion analysis device 207.

The embodiments described above relate specifically to solid collision surface shapes to induce dissolution of clusters on the surface, but (clusters collide with collision surface 209 to induce dissolution at very high speeds). It will be understood that other shapes can be implemented (provided that).

The embodiments described above result in the generation of gas phase sample ions due to collisions with the collision surface, but additional ionization techniques may be used downstream of the collision surface to generate the sample ions. Is intended.

The embodiments described above collide a mixture of matrix and specimen on a collision surface to atomize the mixture, but it is contemplated that alternative atomization techniques may be used.

Although the present invention has been described with reference to various embodiments, various modifications may be made in shape and detail without departing from the scope of the invention as described in the appended claims. That will be understood by those skilled in the art.

Claims (18)

A method of mass spectrometry,
Providing a sample by using the first device to produce an aerosol, smoke or vapor from the target to be analyzed.
Feeding the aerosol, smoke or vapor with the matrix compound such that the sample is diluted by the matrix, dissolved in the matrix, or forms a first cluster in the matrix.
Colliding the first cluster or first droplet of the diluted or dissolved sample against the collision surface placed in the vacuum chamber of the mass spectrometer so as to generate multiple sample ions. including bets, the method of mass spectrometry. The matrix comprises (i) 50-100 μl / min, (ii) 100-150 μl / min, (iii) 150-200 μl / min, (iv) 200-250 μl / min, (v) 250-300 μl / min, (v) vi) 300-350 μl / min, (vii) 350-400 μl / min, (viii) 400-450 μl / min, (ix) 450-500 μl / min, (x) 500-550 μl / min, (xi) 550-600 μl / Min, (xii) 600-650 μl / min, (xiii) 650-700 μl / min, (xiv) 700-750 μl / min, (xv) 750-800 μl / min, (xvi) 800-850 μl / min, (xvii) ) 850 to 900 μl / min, (xviii) 900 to 950 μl / min, (xx) 950 to 1000 μl / min, (xx) 50 μl / min to 1 ml / min, (xxii) 100 to 800 μl / min, (xxiii) 150 to 600 [mu] l / min, and (xxiii) at 200~400μl / min selected flow rate from the group consisting of, the aerosol is supplied to the smoke or vapors, the method of mass spectrometry as claimed in claim 1. Supplying the matrix compound to the aerosol, smoke or vapor
With the matrix molecule in the gas phase, the matrix molecule is supplied to the sample, and the matrix molecule is mixed with the sample.
1 or claim 1, wherein the matrix molecule is in the form of an aerosol, vapor, or solid, the matrix molecule is fed to the sample, and the matrix molecule is mixed with the sample. the method of mass spectrometry as claimed in 2. The mass spectrometric according to any one of claims 1-3, wherein the step of using the first device to produce an aerosol, smoke or vapor from the target further comprises irradiating the target with a laser. Analysis method. The first device includes (i) rapid evaporation ionization mass analysis (“REIMS”) ion source, (ii) desorption electrospray ionization (“DESI”) ionization source, and (iii) laser desorption ionization (“LDI”). ) Ion source, (iv) thermal desorption ion source, (v) laser diode thermal desorption (“LDTD”) ion source, (vi) desorption electrospray focused (“DEFI”) ion source, (vii) dielectric Barrier discharge ("DBD") plasma ion source, (viii) atmospheric solid-state analysis probe ("ASAP") ion source, (ix) ultrasonic-assisted spray ionization ion source, (x) simple atmospheric sonic spray ionization ("EASI") ) Ion source, (xi) desorption atmospheric pressure photoionization ("DAPPI") ion source, (xii) paper spray ("PS") ion source, (xiii) jet desorption ionization ("JeDI") ion source, ( xiv) touch spray (“TS”) ion source, (xv) nanoDISI ion source, (xvi) laser ablation electrospray (“LAESI”) ion source, (xvi) real-time direct analysis (“DART”) ionization source, (xvi) xviii) Probe Electrospray Ionization (“PESI”) Ion Source, (xx) Solid Probe Assisted Electrospray Ionization (“SPA-ESI”) Ion Source, (xx) Cavitron Ultrasonic Surgical Aspiration (“CUSA”) Device, ( xxx) Hybrid CUSA diathermy device, (xxiii) focused or unfocused electrospray ionization device, (xxiii) hybrid focused or unfocused electrospray and diathermy device, (xxiv) microwave resonator, (xxv) pulse plasma RF dissociation device , (Xxxvi) Argon Plasma Coagulation Equipment, (xxxvi) Hybrid Pulse Plasma RF Dissociation and Argon Plasma Coagulation Equipment, (xxvii) Hybrid Pulse Plasma RF Dissociation and JeDI Equipment, (xxxvii) Surgical Water / Physiological Salt Water Jet Equipment, (xxx) A part of a device selected from the group consisting of a hybrid electrospray and argon plasma coagulation device and a (xxx) hybrid argon plasma coagulation and water / physiological saline jet device, or an ion source, which comprises or forms a claim. the method of mass spectrometry as claimed in any one of items 1-3. It said matrix compound comprises a proton matrix solvent, mass fraction method analysis according to any one of claims 1 to 5. The matrix comprises (i) a solvent for a sample, (ii) an organic solvent, (iii) a volatile compound, (iv) a polar molecule, (v) water, (vi) one or more alcohols, (vii). Methanol, (viii) ethanol, (ix) isopranol, (x) acetone, (xi) acetonitrile, (xii) 1-butanol, (xiii) tetrahydrofuran, (xiv) ethyl acetate, (xv) ethylene glycol, (xvi) A claim selected from the group consisting of dimethyl sulfoxide, (xvii) aldehyde, (xvii) ketone, (xiv) non-polar molecule, (xx) hexane, (xxi) chloroform, (xxxi) butanol, and (xxiii) propanol. the method of mass spectrometry as claimed in any one of items 1-6. Further comprising a weight fraction method analysis according to any one of claims 1 to 7 using the pressure difference the first cluster or the first liquid droplet in order to accelerate the impact surface. The method of mass spectrometry according to any one of claims 1 to 8, further comprising heating the collision surface. A quality amount diffractometer,
A first device that produces an aerosol, smoke or vapor from a target to be analyzed and provides a sample.
A device that feeds a matrix compound to an aerosol, smoke or vapor such that the sample is diluted by a matrix, dissolved in the matrix, or forms a first cluster in the matrix.
The first cluster or first liquid of the diluted or dissolved sample of the collision surface placed in the vacuum chamber of the mass spectrometer so as to generate multiple sample ions during use. and a collision surface impinging drops, mass fraction diffractometer. The matrix comprises (i) 50-100 μl / min, (ii) 100-150 μl / min, (iii) 150-200 μl / min, (iv) 200-250 μl / min, (v) 250-300 μl / min, (v) vi) 300-350 μl / min, (vii) 350-400 μl / min, (viii) 400-450 μl / min, (ix) 450-500 μl / min, (x) 500-550 μl / min, (xi) 550-600 μl / Min, (xii) 600-650 μl / min, (xiii) 650-700 μl / min, (xiv) 700-750 μl / min, (xv) 750-800 μl / min, (xvi) 800-850 μl / min, (xvii) ) 850 to 900 μl / min, (xviii) 900 to 950 μl / min, (xx) 950 to 1000 μl / min, (xx) 50 μl / min to 1 ml / min, (xxii) 100 to 800 μl / min, (xxiii) 150 to 600 [mu] l / min, and (xxiii) at a flow rate selected from the group consisting of 200~400μl / min, the aerosol is supplied to the smoke or vapors, mass fraction analyzers according to claim 10. A device that supplies the matrix compound to the aerosol, smoke or vapor
With the matrix molecule in the gas phase, the matrix molecule is supplied to the sample, and the matrix molecule is mixed with the sample.
With the matrix molecules in the form of an aerosol, vapor, or solid, it was configured to feed the matrix molecules to the sample and mix the matrix molecules with the sample at least one of them. , mass fraction analyzers according to claim 10 or 11. It said first device comprises a laser, mass fraction analyzers according to any one of claims 10-12. The first device includes (i) rapid evaporation ionization mass analysis (“REIMS”) ion source, (ii) desorption electrospray ionization (“DESI”) ionization source, and (iii) laser desorption ionization (“LDI”). ) Ion source, (iv) thermal desorption ion source, (v) laser diode thermal desorption (“LDTD”) ion source, (vi) desorption electrospray focused (“DEFI”) ion source, (vii) dielectric Barrier discharge ("DBD") plasma ion source, (viii) atmospheric solid-state analysis probe ("ASAP") ion source, (ix) ultrasonic-assisted spray ionization ion source, (x) simple atmospheric sonic spray ionization ("EASI") ) Ion source, (xi) desorption atmospheric pressure photoionization ("DAPPI") ion source, (xii) paper spray ("PS") ion source, (xiii) jet desorption ionization ("JeDI") ion source, ( xiv) touch spray (“TS”) ion source, (xv) nanoDISI ion source, (xvi) laser ablation electrospray (“LAESI”) ion source, (xvi) real-time direct analysis (“DART”) ionization source, (xvi) xviii) Probe Electrospray Ionization (“PESI”) Ion Source, (xx) Solid Probe Assisted Electrospray Ionization (“SPA-ESI”) Ion Source, (xx) Cavitron Ultrasonic Surgical Aspiration (“CUSA”) Device, ( xxx) Hybrid CUSA diathermy device, (xxiii) focused or unfocused electrospray ionization device, (xxiii) hybrid focused or unfocused electrospray and diathermy device, (xxiv) microwave resonator, (xxv) pulse plasma RF dissociation device , (Xxvi) Argon Plasma Coagulation Equipment, (xxxvi) Hybrid Pulse Plasma RF Dissociation and Argon Plasma Coagulation Equipment, (xxxvii) Hybrid Pulse Plasma RF Dissociation and JeDI Equipment, (xxxvii) Surgical Water / Physiological Salt Water Jet Equipment, (xxx) A part of a device selected from the group consisting of a hybrid electrospray and argon plasma coagulation device and a (xxx) hybrid argon plasma coagulation and water / physiological saline jet device, or an ion source, which comprises or forms a claim. mass fraction analyzers according to any one of clauses 10-12. It said matrix compound comprises a proton matrix solvent, mass fraction analyzers according to any one of claims 10-14. The matrix comprises (i) a solvent for a sample, (ii) an organic solvent, (iii) a volatile compound, (iv) a polar molecule, (v) water, (vi) one or more alcohols, (vii). Methanol, (viii) ethanol, (ix) isopranol, (x) acetone, (xi) acetonitrile, (xii) 1-butanol, (xiii) tetrahydrofuran, (xiv) ethyl acetate, (xv) ethylene glycol, (xvi) A claim selected from the group consisting of dimethyl sulfoxide, (xvii) aldehyde, (xvii) ketone, (xiv) non-polar molecule, (xx) hexane, (xxx) chloroform, (xxxi) butanol, and (xxiii) propanol. mass fraction analyzers according to any one of clauses 10-14. A pressure difference is created between the first and second regions in order to accelerate the first cluster or first droplet between the first region and the second region to the collision surface. mass fraction analyzers of configured, according to any one of claims 10-16 as. The mass spectrometer according to any one of claims 10 to 17, further comprising a heater for heating the collision surface.

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